Apparatus and method for surgical stereotactic procedures

An apparatus and method are disclosed for displaying a path between a selected target and selected points on a patient's skull and for guiding surgical instruments along any selected path. The system is comprised of an image display system, an articulated arm and probe, and a stereotactic system. The sub-systems are coupled to one another so that the articulated probe may be used to select patient fiducial points that correspond to selected image fiducial points. Using these points, the image display system coregisters the external locations to the displayed images so that the probe condition may be displayed with the displayed images. The system further permits the identification of a selected target within a patient's brain and to project a path from the external position to the target prior to the performance of a craniotomy. After evaluation of the path, a surgeon may lock the stereotactic system in place to preserve a selected surgical path and to guide instruments along that path.

Acoustic perfusion devices

Acoustic perfusion devices for separating biological cells from other material in a fluid medium are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid medium is being drawn.

ACOUSTIC PERFUSION DEVICES

Acoustic perfusion devices for separating biological cells from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn. 1. An acoustic perfusion device , comprising:an acoustic chamber;an inlet port and an inlet flow path leading from the inlet port to the acoustic chamber;an outlet port and an outlet flow path leading from the acoustic chamber to the outlet port for recirculating a fluid mixture;at least one collection port for collecting harvest fluid from the fluid mixture; andat least one ultrasonic transducer and at least one reflector opposite the at least one ultrasonic transducer, wherein the at least one ultrasonic transducer includes a piezoelectric material driven by a voltage signal to create a multi-dimensional acoustic standing wave across a collection flow path leading from the acoustic chamber to the at least one collection port.2. The device of claim 1 , wherein the acoustic chamber is below the at least one collection port.3. The device of claim 1 , wherein the inlet flow path is shaped to generate a tangential flow path below an acoustic field generated by the acoustic standing wave.4. The device of claim 1 , wherein a pressure rise and an acoustic radiation force on cells are generated at an upstream interface region of the acoustic standing wave to clarify fluid passing through the acoustic standing wave.5. The device of claim 1 , wherein the at least one reflector is made of a transparent material.6. The device of claim 1 , wherein the outlet port is below the inlet port.7. The device of claim 1 , having a total of two or more ...

SYSTEMS AND METHODS FOR ADAPTIVE CONTROL OF WIRE PREHEATING

A welding-type system includes a power supply configured to control preheating of an electrode wire. A controller is configured to receive a plurality of power values corresponding to a power output of the power supply and calculate an arc power value corresponding to an arc condition at the preheated electrode wire based on a rate of change of the plurality of power values. A target power output value is determined based on the calculated arc power value, and the power output is adjusted based on the determined target power value. 1. A welding-type system , comprising:a power supply configured to preheat an electrode wire; and receive a plurality of power values corresponding to a power output of the power supply;', 'calculate an arc power value corresponding to an arc condition at the preheated electrode wire based on a rate of change of the plurality of power values;', 'determine a target power output value based on the calculated arc power value; and', 'adjust the power output based on the determined target power value., 'a controller configured to2. The system as defined in claim 1 , wherein the power output is regulated between a high threshold value and a low threshold value about the target power output value.3. The system as defined in claim 2 , wherein the controller is configured to:compare a first power value of the plurality of power values to the high threshold power value and the low threshold power value; andadjust the power output based on the comparison.4. The system as defined in claim 2 , wherein the controller is configured to:raise the power output if the first power output is below the low threshold power value; andto lower the power output if the first power output is above the high threshold power value.5. The system as defined in claim 2 , further comprising a wire feeder claim 2 , the controller further configured to adjust a wire feed speed based on a comparison of the first power value to the target power output value and the high and ...

MULTI-AMPLITUDE MODULAR LABELED COMPOUNDS

Sets of compounds bearing detectably different groups of labels are provided. Typically, different compounds bear different numbers of a single type of label and are thus distinguishable by the amplitude of signal produced by the label. The compounds are assembled from label components and protein cores to facilitate modular production of the compounds. In compounds containing two or more proteins, the proteins are typically covalently linked. Useful sets of compounds include sets of labeled nucleotide analogs, particularly dye-label nucleotide analogs that include tetravalent biotin-binding protein cores. 1. A set of dye-labeled nucleotide analogs , the set comprising:a first labeled nucleotide analog comprising i) one or more tetravalent biotin-binding proteins, ii) one or more first nucleotide components bound to the tetravalent biotin-binding proteins, and iii) one or more dye components bound to the tetravalent biotin-binding proteins, each dye component comprising one or more dye moieties; anda second labeled nucleotide analog comprising i) two or more covalently linked tetravalent biotin-binding proteins, ii) one or more second nucleotide components bound to the tetravalent biotin-binding proteins, and iii) two or more dye components bound to the tetravalent biotin-binding proteins, each dye component comprising one or more dye moieties;wherein the total number of dye moieties in the second labeled nucleotide analog is greater than that in the first labeled nucleotide analog.2. (canceled)3. The set of claim 1 , wherein each component is bound to the tetravalent biotin-binding protein through a biotin moiety.4. The set of claim 1 , wherein each component comprises a bis-biotin moiety bound to two biotin binding sites on the tetravalent biotin-binding protein.5. The set of claim 1 , comprising a third labeled nucleotide analog comprising i) two or more covalently linked tetravalent biotin-binding proteins claim 1 , ii) one or more third nucleotide components ...

Recombinant polymerases for incorporation of protein shield nucleotide analogs

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

METHODS AND COMPOSITIONS FOR LOADING OF POLYMERASE COMPLEXES

The present disclosure provides methods, compositions, and systems for distributing polymerase compositions into array regions. In particular, the described methods, compositions, and systems utilize density differentials and/or additives to increase efficiency in the distribution of polymerase compositions to a surface as compared to methods utilizing only diffusion control. 195-. (canceled)97. The method of claim 96 , wherein the molecules comprise nucleic acid.98. The method of claim 96 , wherein the molecules comprise polymerase enzymes.99. The method of claim 96 , wherein the molecules comprise helicases.100. The method of claim 96 , wherein the molecules comprise complexes of (i) polymerase enzymes and nucleic acid templates or (ii) helicases and nucleic acid templates.101. The method of claim 96 , wherein the spike second solution comprises a neutral and hydrophilic polysaccharide.102. The method of claim 96 , wherein the spike second solution comprises a highly branched claim 96 , high-mass polysaccharide.103. The method of claim 96 , wherein the second solution comprises a volume excluding buffer reagent.104. The method of claim 96 , wherein the spike second solution comprises an additive selected from the group consisting of dextran claim 96 , aminodextran claim 96 , dextrin claim 96 , cluster dextrin claim 96 , Ficoll claim 96 , polyetheylene glycol claim 96 , sucrose claim 96 , DMSO claim 96 , glycerol claim 96 , and pullulan.105. The method of claim 96 , wherein the second solution comprises Ficoll.106. The method of claim 96 , wherein the distributing occurs at about a 2 to about a 25-fold faster rate as compared to distributing without the second solution.107. The method of claim 96 , wherein the distributing occurs at about a 5 to about a 20-fold faster rate as compared to distributing without the second solution.108. The method of claim 96 , wherein the distributing occurs at about a 10 to about a 15-fold faster rate as compared to distributing ...

Recombinant polymerases for incorporation of protein shield nucleotide analogs

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Labeled nucleotide analogs having protein shields

Compositions, methods, and systems are provided for nucleotide analogs comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Nucleotide analogs of the invention have a protein shield between the dye moieties and nucleotide moieties of the analog. The nucleotide analogs have two avidin proteins connected by a central dye component, and each avidin protein is further connected to a nucleotide component. The protein can prevent the direct interaction of the dye moiety with the enzyme carrying out nucleotide synthesis preventing photodamage to the enzyme. The nucleotide analogs of the invention can have multiple dyes and multiple nucleotide moieties.

Acoustic perfusion devices

Methods are disclosed for separating beads and cells from a host fluid. The method includes flowing a mixture containing the host fluid, the beads, and the cells through an acoustophoretic device having an ultrasonic transducer including a piezoelectric material driven by a drive signal to create a multi-dimensional acoustic standing wave. A drive signal is sent to drive the at least one ultrasonic transducer to create the multi-dimensional acoustic standing wave. A recirculating fluid stream having a tangential flow path is located substantially tangential to the standing wave and separated therefrom by an interface region. A portion of the cells pass through the standing wave, and the beads are held back from the standing wave in the recirculating fluid stream at the interface region. Also disclosed is an acoustophoretic device having a coolant inlet adapted to permit the ingress of a cooling fluid into the device for cooling the transducer.

DELAYING REAL-TIME SEQUENCING

Methods, compositions, and systems are provided that allow for reliable sequencing of the initial sequence region of a sequence of interest. The methods of the invention allow for more reliable barcoding of subpopulations of nucleic acids to be sequenced.

ACOUSTIC PERFUSION DEVICES

Acoustic perfusion devices for separating biological cells from other material in a fluid medium are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid medium is being drawn. 1. An acoustic perfusion device , comprising:an acoustic chamber; an inlet port, an inlet flow path leading from the inlet port to the acoustic chamber;an outlet port for recirculating a fluid/cell mixture;at least one collection port for collecting harvest fluid from the fluid/cell mixture; andat least one ultrasonic transducer and at least one reflector opposite the at least one ultrasonic transducer, wherein the at least one ultrasonic transducer includes a piezoelectric material driven by a voltage signal to create a multi-dimensional acoustic standing wave across a collection flow path leading from the acoustic chamber to the at least one collection port.2. The device of claim 1 , wherein the acoustic chamber is below the at least one collection port.3. The device of claim 1 , wherein the inlet flow path is shaped to generate a tangential flow path below an acoustic field generated by the acoustic standing wave.4. The device of claim 1 , wherein a pressure rise and an acoustic radiation force on cells are generated at an edge of the acoustic standing wave to clarify fluid passing through the acoustic standing wave.5. The device of claim 1 , wherein the at least one ultrasonic transducer is mounted in a rear wall or a front wall of the device.6. The device of claim 1 , wherein the at least one reflector is made of a transparent material.7. The device of claim 1 , wherein the outlet port is below the inlet port.8. The device of claim 1 , ...

ACOUSTIC PERFUSION DEVICES

Methods are disclosed for separating beads and cells from a host fluid. The method includes flowing a mixture containing the host fluid, the beads, and the cells through an acoustophoretic device having an ultrasonic transducer including a piezoelectric material driven by a drive signal to create a multi-dimensional acoustic standing wave. A drive signal is sent to drive the at least one ultrasonic transducer to create the multi-dimensional acoustic standing wave. A recirculating fluid stream having a tangential flow path is located substantially tangential to the standing wave and separated therefrom by an interface region. A portion of the cells pass through the standing wave, and the beads are held back from the standing wave in the recirculating fluid stream at the interface region. Also disclosed is an acoustophoretic device having a coolant inlet adapted to permit the ingress of a cooling fluid into the device for cooling the transducer. 1. An acoustic bead retention device , comprising:a chamber;at least one ultrasonic transducer coupled to the chamber;the at least one ultrasonic transducer configured to be excited to generate a multi-dimensional acoustic standing wave that is configured to retain beads and pass fluid in which the beads are entrained.2. The device of claim 1 , further comprising a recirculation path communicating with the acoustic chamber.3. The device of claim 1 , wherein the multi-dimensional acoustic standing wave is further configured to generate a pressure rise and an acoustic radiation force on the beads at an interface region of the multi-dimensional acoustic standing wave.4. The device of claim 3 , further comprising a locale adjacent to the interface region where the bead concentration is increased.5. The device of claim 1 , further comprising a locale in the multi-dimensional acoustic standing wave where the bead concentration is increased.6. The device of claim 1 , wherein the multi-dimensional acoustic standing wave is further ...

Methods, systems, and reagents for direct RNA sequencing

Provided are compositions comprising recombinant polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for sequencing RNA or RNA/DNA templates. Polymerases that topologically encircle the template nucleic acid are provided. Also provided are methods of using such polymerases to make a DNA or to sequence a template comprising RNA.

DEPLOYABLE KIOSK FOR PROVIDING DATA INPUT AND VERIFICATION

According to exemplary embodiments, a secure kiosk for receiving, transmitted, securing, and verifying data may be shown and described. For example, the kiosk may be fully automated. The kiosk may include a display, a tablet computer, a card reader, a printer, and a network link. For example, a kiosk may be provided and may be fully automated. The kiosk may accept data inputs in a variety of manners. The inputted data may then be transmitted, in real time, to a first server, where the information can be displayed and stored. Additionally, the information, at predetermined times, may be transmitted to a number of other servers which may receive, compare, and validate information. Further, a hierarchical data transmission system may be utilized such that, when information is verified or indicated as unverified at any of the other servers, instructions may be sent to at least the first server and the kiosk. 1. A deployable kiosk comprising a display , a card reader , a printer , a tablet having a processor , and a network link , the deployable kiosk operable to:receive user information comprising at least the name of a user and a location of the user;transmit the user information to a first server;display, on a graphical user interface of a user terminal, the user information;transmit the user information to at least one second server, the at least one second server being a database server storing identity verification data;receive a verification message from the at least one second server, the verification message comprising a verification of the authenticity of the user information of the user;determine the geographic eligibility of the user to be provided a geographically-limited good or service;determine location data of the user terminal;determine, from the location data of the user terminal, whether the user terminal is within an eligible area to be provided a geographically-limited good or service, and whether the user terminal is in its intended location; ...

MODIFIED BIOTIN-BINDING PROTEINS FOR IMMOBILIZATION

Compositions comprising covalently modified and mutated biotin-binding proteins, particularly biotin-binding proteins having a negative charge at physiological pH, are provided. Methods of producing such proteins are also provided, as are methods of immobilizing, sequencing, and making nucleic acids employing such proteins. 1. (canceled)2. The system of claim 152 , wherein the biotin-binding protein is a tetravalent biotin-binding protein.3. The system of claim 152 , wherein the biotin-binding protein is streptavidin.4. (canceled)5. The system of claim 152 , wherein the biotin-binding protein comprises 12 or more covalently attached sulfonate moieties.6. The system of claim 152 , wherein the biotin-binding protein comprises 24 or more covalently attached sulfonate moieties.7. The system of claim 152 , wherein the biotin-binding protein comprises 30 or more covalently attached sulfonate moieties.8. The system of claim 152 , wherein the biotin-binding protein comprises 45 or more covalently attached sulfonate moieties.9. (canceled)10. The system of claim 152 , wherein the biotin-binding protein comprises one or more covalently attached 3 claim 152 ,4 claim 152 ,5-tris(3-sulfopropoxy)benzoyl moieties.11. The system of claim 152 , wherein the biotin-binding protein comprises four or more covalently attached 3 claim 152 ,4 claim 152 ,5-tris(3-sulfopropoxy)benzoyl moieties.12. The system of claim 152 , wherein the biotin-binding protein comprises 10 or more covalently attached 3 claim 152 ,4 claim 152 ,5-tris(3-sulfopropoxy)benzoyl moieties.13. The system of claim 152 , wherein the biotin-binding protein comprises 15 or more covalently attached 3 claim 152 ,4 claim 152 ,5-tris(3-sulfopropoxy)benzoyl moieties.14. (canceled)15. The system of claim 152 , wherein the biotin-binding protein is a tetravalent biotin-binding protein comprising 45 or more covalently attached sulfonate moieties.16. The system of claim 15 , wherein the biotin-binding protein is streptavidin.17. The ...

Wire feed limiter

A wire conveying mechanism, preferably for a welding, cladding or additive manufacturing apparatus, with a slip clutch mechanism connectable to a motor.

DELAYING REAL-TIME SEQUENCING

Methods, compositions, and systems are provided that allow for reliable sequencing of the initial sequence region of a sequence of interest. The methods of the invention allow for more reliable barcoding of subpopulations of nucleic acids to be sequenced. 170-. (canceled)7130. A composition comprising a plurality of nucleic acid sequencing templates , wherein each of the plurality nucleic acid sequencing templates comprises an adaptor region and an insert region , wherein the adaptor region comprises a sequencing initiation site and a runway region and the insert region comprises a sequence of interest , wherein the runway region is between the sequencing initiation site and the insert region , and wherein the runway region has a length that delays sequencing the insert region for seconds or more in a nucleic acid sequencing reaction initiated from the sequencing initiation site.72. The composition of claim 71 , wherein the runway region has a length that delays sequencing the insert region is greater than about 30 seconds.73. The composition of claim 72 , wherein the length of the runway region is 200 nucleotides or greater.74. The composition of claim 73 , wherein the length of the runway region is between 200 and 2 claim 73 ,000 nucleotides.75. The composition of claim 71 , wherein the runway region comprises at least one modified base that is absent from the insert region.76. The composition of claim 71 , wherein the runway region in each of the plurality of nucleic acid sequencing templates has substantially the same sequence.77. The composition of claim 71 , wherein each of the plurality of nucleic acid sequencing templates is linear.78. The composition of claim 77 , wherein each of the plurality of nucleic acid sequencing templates comprises a single strand region on at least one end claim 77 , wherein the single strand region comprises the sequencing initiation site.79. The composition of claim 78 , wherein the sequencing initiation site is an initiation ...

RECOMBINANT POLYMERASES FOR IMPROVED SINGLE MOLECULE SEQUENCING

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Bis-biotinylation tags

Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants.

Acoustic perfusion devices

Acoustic perfusion devices for separating biological cells from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn.

RECOMBINANT POLYMERASES WITH INCREASED PHOTOTOLERANCE

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 120-. (canceled)21. A composition comprising a ϕ29-type (phi29-type) recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position S10 , an amino acid substitution at position E244 , an amino acid substitution at position A444 , an amino acid substitution at position I460 , a T231S substitution , a K361R substitution , a T573E substitution , a T573G substitution , and a T573S substitution , wherein identification of positions is relative to SEQ ID NO:1 , and wherein said polymerase exhibits polymerase activity.22. The composition of claim 21 , wherein the recombinant polymerase comprises one or more mutation selected from the group consisting of an S10V substitution claim 21 , an E244K substitution claim 21 , an E244Q substitution claim 21 , an E244R substitution claim 21 , an A444G substitution claim 21 , an A444L substitution claim 21 , an A444S substitution claim 21 , an A444V substitution claim 21 , an 1460F substitution claim 21 , ...

LASER WELDING ENCLOSURE

Systems and methods are disclosed for an enclosure providing a sealed environment. The enclosure employs movable paneling that seals an area for a laser welding cell, while allowing access for an operator to load a workpiece in and unload the workpiece from a welding cell. Barriers are arranged on the enclosure, such that in a closed configuration activation of a pneumatic tube forces the barriers to extend toward a rotated paneling, such that the edges of the enclosure contact or overlap with the edges of the paneling to result in a light-tight seal. Presence of the fluid increases the pressure in the tube such that an external boundary of the tube expands, forcing the barrier toward the edges of the paneling, thereby closing the gap and providing a light-tight enclosure.

ACOUSTIC PERFUSION DEVICES

Methods are disclosed for separating beads and cells from a host fluid. The method includes flowing a mixture containing the host fluid, the beads, and the cells through an acoustophoretic device having an ultrasonic transducer including a piezoelectric material driven by a drive signal to create a multi-dimensional acoustic standing wave. A drive signal is sent to drive the at least one ultrasonic transducer to create the multi-dimensional acoustic standing wave. A recirculating fluid stream having a tangential flow path is located substantially tangential to the standing wave and separated therefrom by an interface region. A portion of the cells pass through the standing wave, and the beads are held back from the standing wave in the recirculating fluid stream at the interface region. Also disclosed is an acoustophoretic device having a coolant inlet adapted to permit the ingress of a cooling fluid into the device for cooling the transducer. 1. A method for separating a desired target material from a host fluid , the method comprising: a flow chamber including at least one inlet and at least one outlet;', 'at least one ultrasonic transducer coupled to the flow chamber, the at least one ultrasonic transducer including a piezoelectric material; and', 'a reflector located opposite from the at least one ultrasonic transducer; and, 'flowing a mixture containing the host fluid, beads, and the target material through an acoustophoretic device, the acoustophoretic device comprisingdriving the at least one ultrasonic transducer to create a multi-dimensional acoustic standing wave in the flow chamber;wherein the beads attach to the target material and are held back by the multi-dimensional acoustic standing wave; and wherein the host fluid passes through the multi-dimensional acoustic standing wave.2. The method of claim 1 , wherein the at least one outlet includes (i) a permeate outlet through which the host fluid exits the flow chamber; and (ii) a concentrate outlet ...

Decorative illumination apparatus

A novel illumination apparatus that may be formed from any object of choice, such as a natural rock or the like. The apparatus is of simple construction and allows a user to easily adjust the amount of light to be emitted there from. Or the user may direct the light in a preferred direction, and optional accessories are also provided, such as the apparatus may include, a motion detector, speakers, a cup-holder, a secret compartment, etc.

ACOUSTIC PERFUSION DEVICES

Acoustic perfusion devices for separating biological cells from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn. 1. An acoustic perfusion device , comprising:an acoustic chamber;at least one ultrasonic transducer coupled to the acoustic chamber;at least one reflector opposite the at least one ultrasonic transducer across the acoustic chamber;the at least one ultrasonic transducer configured to be excited to generate an acoustic standing wave with the at least one reflector;a flow path arranged in the acoustic chamber to bring a fluid and particle mixture proximate to the acoustic standing wave, such that a perfusate turbidity is reduced by about 50% or greater.2. The device of claim 1 , wherein the flow path is shaped to generate a tangential flow path proximate to the acoustic standing wave.3. The device of claim 1 , wherein a pressure rise and an acoustic radiation force on cells are generated at an upstream interface region of the acoustic standing wave to clarify fluid passing through the acoustic standing wave.4. The device of claim 1 , wherein the at least one reflector is made of a transparent material.5. The device of claim 1 , wherein the at least one ultrasonic transducer further comprises two or more ultrasonic transducers.6. The device of claim 5 , where the at least one reflector is located between the two or more ultrasonic transducers.7. The device of claim 1 , wherein the acoustic standing wave generates an acoustic radiation force with an axial force component and a lateral force component that are of the same order of magnitude.8. ...

Acoustic perfusion devices

Acoustic perfusion devices for separating biological cells from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn.

DISPENSER CONTROL SYSTEMS AND METHODS

A method of operating a dispensing system having a material delivery cycle. In some embodiments, the material delivery cycle includes supplying water to a receptacle, performing an operation intended to release a material into the water, and delivering the material to a downstream component. The first step of the method is to initiate the material delivery cycle. Next, a conductivity proximate to the receptacle is monitored. Additionally, one or more error conditions are identified during the material delivery cycle based at least partially on the monitored conductivity.

BIS-BIOTINYLATION TAGS

Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants. 131-. (canceled)33. The compound of claim 32 , wherein Lis a linking group comprising a negatively charged group and Lis a linking group comprising a negatively charged group.34. The compound of claim 33 , wherein Land Lcomprise a sulfonic acid moiety.35. The compound of claim 33 , wherein Land Lcomprise a methylsulfonic acid group.36. The compound of claim 33 , wherein Land Leach comprise three sulfonic acid moieties.38. A method of preparing a surface claim 32 , the method comprising coating the surface with the compound of claim 32 , wherein reaction of the compound with the surface covalently couples the two biotin groups to the surface.3941-. (canceled)42. The method of claim 38 , wherein the surface comprises the bottom surface of at least one nanoscale well.43. The method of claim 38 , comprising binding a tetravalent biotin-binding agent to the two biotin groups.44. The method of claim 43 , comprising binding a molecule or molecular complex of interest to the tetravalent biotin-binding agent.45. The method of claim 44 , wherein binding the molecule or molecular complex of interest to the tetravalent biotin-binding agent is performed before binding the tetravalent biotin-binding agent to the two biotin groups.46. The method of claim 44 , wherein a bis-biotin tag is covalently coupled to the molecule or molecular complex of interest claim 44 , and wherein the bis-biotin tag is bound to the tetravalent biotin-binding agent.47. The method of claim 44 , wherein the molecule of interest is an enzyme.48. ...

BIS-BIOTINYLATION TAGS

Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants. 1. A solid support whose surface comprises multiple bis-biotin moieties , each bis-biotin moiety having two covalently linked biotin moieties.218-. (canceled)20. The compound of claim 19 , wherein F is a carboxylic acid group or an amino group.21. The compound of claim 19 , wherein F is an azide group or an alkyne group.22. The compound of claim 19 , wherein Land Leach comprise a methylsulfonic acid group.23. The compound of claim 19 , wherein Land Leach comprise three sulfonic acid moieties.24. The compound of claim 19 , wherein Land Leach comprise a 3 claim 19 ,4 claim 19 ,5-tris(3-sulfopropoxy)benzoic acid group.25. The compound of claim 19 , wherein Land Lare identical groups.26. The compound of claim 19 , wherein Land Lcomprise different groups.27. The compound of claim 19 , wherein C comprises a six-membered ring.28. The compound of claim 27 , wherein C comprises a tri-substituted aromatic six-membered ring.29. The compound of claim 27 , wherein C comprises a hi-substituted triazine.30. The compound of claim 27 , wherein C comprises a hi-substituted cyclohexane ring.3337-. (canceled)38. A method of preparing a surface claim 27 , the method comprising coating the surface with a compound that comprises a bis-biotin moiety claim 27 , each bis-biotin moiety having two covalently linked biotin moieties.3953-. (canceled)54. The compound of claim 19 , wherein Land Leach comprise a sulfonic acid moiety. This application is a non-provisional utility patent application claiming priority to and benefit of ...

Methods and compositions for enriching compositions for polymerase enzyme complexes

The present invention provides methods, compositions, and systems for enriching compositions for polymerase enzyme complexes. In particular, the methods, compositions, and systems of the present invention remove free polymerases from the compositions using one or more purification steps, including protease treatment, thus enriching the compositions for polymerases complexed with a template nucleic acid.

DELAYING REAL-TIME SEQUENCING

Methods, compositions, and systems are provided that allow for reliable sequencing of the initial sequence region of a sequence of interest. The methods of the invention allow for more reliable barcoding of subpopulations of nucleic acids to be sequenced.

LASER WELDING, CLADDING, AND/OR ADDITIVE MANUFACTURING SYSTEMS AND METHODS OF LASER WELDING, CLADDING, AND/OR ADDITIVE MANUFACTURING

Systems and methods of a laser welding device are disclosed. The laser welding device includes a laser generator configured to generate welding-type lasing power. A lens focuses the welding-type lasing power at a focal point on a workpiece to generate a puddle during a welding-type operation. A wire feeder is configured to feed wire to the puddle generated by the laser generator. A laser scanner controls the lens to move the focal point of the welding-type lasing power in multiple dimensions over the workpiece during the welding-type operation. In some examples, the feed wire is used in an additive manufacturing process.

ALUMINUM METAL MATRIX COMPOSITE SHEATHS FOR WIRE ELECTRODES

The present disclosure relates to tubular welding electrodes that have a metallic sheath surrounding a granular core, wherein the metallic sheath comprises a metal matrix composite (MMC) that includes a ceramic material and aluminum or an aluminum alloy. The ceramic material may be in the form of microparticles or nanoparticles. The present disclosure also relates to method for making such tubular welding electrodes. 1. A tubular welding electrode comprising:a metallic sheath surrounding a granular core,wherein the metallic sheath comprises a metal matrix composite (MMC) that comprises a ceramic material and aluminum or an aluminum alloy.2. The tubular welding electrode of claim 1 , wherein the ceramic material comprises up to 20% by weight percent of the metallic sheath.3. The tubular welding electrode of claim 1 , wherein the ceramic material is chosen from the group consisting of: alumina (AlO) claim 1 , boron carbide (BC) claim 1 , carbon nanotubes (CNT) claim 1 , titanium dioxide (TiO) claim 1 , silicon carbide (SiC) claim 1 , tungsten carbide (WC) claim 1 , silicon nitride (SiN) claim 1 , aluminum nitride (AlN) claim 1 , titanium carbide (TiC) claim 1 , or silica (SiO).4. The tubular welding electrode of claim 1 , wherein the ceramic material is in the form of microparticles.5. The tubular welding electrode of claim 1 , wherein the ceramic material is in the form of nanoparticles.6. The tubular welding electrode of claim 1 , wherein the MMC comprises a 4xxx series or a 5xxx series aluminum alloy.7. The tubular welding electrode of claim 1 , wherein the granular core is a granular powder flux fill core.8. The tubular welding electrode of claim 1 , wherein the granular core is a granular metal core.9. The tubular welding electrode of claim 1 , wherein the granular core comprises a core ceramic material and wherein the core ceramic material comprises up to 4% by weight percent of the tubular welding electrode.10. A method for producing a tubular welding electrode ...

Recombinant polymerases for improved single molecule sequencing

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Recombinant Polymerases for Incorporation of Protein Shield Nucleotide Analogs

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 1. A composition comprising:a nucleotide analog, which analog comprises a polypeptide moiety positioned between a dye moiety and a nucleotide component of the analog; anda recombinant DNA polymerase, which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or at least 80% identical to SEQ ID NO:2, and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position Q99, an amino acid substitution at position A134, an amino acid substitution at position K138, an amino acid substitution at position V141, an amino acid substitution at position L142, an amino acid substitution at position A256, an amino acid substitution at position R306, an amino acid substitution at position R308, an amino acid substitution at position K311, an amino acid substitution at position T441, an amino acid substitution at position E466, an amino acid substitution at position D476, an amino acid substitution at position ...

Polymerase enzyme substrates with protein shield

Compositions and methods are provided for nucleotide analogs comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Nucleotide analogs of the invention have a protein shield between the dye moieties and nucleotide moieties of the analog. The protein prevents the direct interaction of the dye moiety with the enzyme carrying out nucleotide synthesis preventing photodamage to the enzyme. The nucleotide analogs of the invention can have multiple dyes and multiple nucleotide moieties.

Metalworking system with force controlled wire feed start operation

A method for controlling a start of a metalworking operation. The method includes detecting an initial contact between a wire being fed from a welding apparatus and a workpiece and, in response to the detection, halting feeding of the wire from the welding apparatus. The method further includes activating a high energy heat source configured to heat a tip of the wire and resuming the feeding of the wire from the welding apparatus when the tip of the wire is heated by the high energy heat source to a plastic state. The feeding of the wire is resumed by measuring a force feedback from the wire contacting the workpiece. An apparatus for implementing the method is also disclosed.

METHODS AND COMPOSITIONS FOR LOADING OF POLYMERASE COMPLEXES

The present disclosure provides methods, compositions, and systems for distributing polymerase compositions into array regions. In particular, the described methods, compositions, and systems utilize density differentials and/or additives to increase efficiency in the distribution of polymerase compositions to a surface as compared to methods utilizing only diffusion control.

WIDE PATH WELDING, CLADDING, ADDITIVE MANUFACTURING

A welding or cladding apparatus in which one or more energy beam emitters are used to generate a wide beam spot transverse to a welding or cladding path, and one or more wide feeders feed wire to the spot to create a wide welding or cladding puddle. 1. A metalworking apparatus , comprising:one or more energy beam emitters arranged to irradiate a workpiece and have one or more spots of high energy incident on the workpiece; andone or more wire feeders configured to feed one or more wires to the one or more spots and which when melted by the energy beam or beams form a welding puddle, each spot having a width in a direction transverse to a direction of a welding path, the cumulative width of the one or more spots being greater than a length of the one or more welding spots in the direction of the welding path.2. The apparatus of claim 1 , comprising a plurality spots continuously arrayed along a straight line.3. The apparatus of claim 2 , wherein the width direction of the one or more spots is orthogonal to the welding path.4. The apparatus of claim 2 , wherein the width direction of the one or more spots is at an oblique angle to the welding path.5. The apparatus of claim 1 , wherein the cumulative width of the spots is 4 mm or greater.6. The apparatus of claim 1 , comprising a single energy beam emitter which emits a wide energy beam or a moving energy beam to create a wide welding puddle.7. The apparatus of claim 1 , comprising a plurality energy beam emitters which emit energy beams of substantially equal cross-section and intensity.8. The apparatus of claim 1 , comprising a plurality energy beam emitters which emit energy beams which different energy intensities.9. The apparatus of claim 1 , wherein each energy beam emitter is a laser.10. The apparatus of claim 5 , wherein an intensity of the energy varies across the spot.11. A metalworking process comprising:irradiating a workpiece with one or more beams of energy and creating one or more spots of high energy ...

Metalworking system with force controlled wire feed start operation

A method for controlling a start of a metalworking operation. The method includes detecting an initial contact between a wire being fed from a welding apparatus and a workpiece and, in response to the detection, halting feeding of the wire from the welding apparatus. The method further includes activating a high energy heat source configured to heat a tip of the wire and resuming the feeding of the wire from the welding apparatus when the tip of the wire is heated by the high energy heat source to a plastic state. The feeding of the wire is resumed by measuring a force feedback from the wire contacting the workpiece. An apparatus for implementing the method is also disclosed.

MULTI-AMPLITUDE MODULAR LABELED COMPOUNDS

Sets of compounds bearing detectably different groups of labels are provided. Typically, different compounds bear different numbers of a single type of label and are thus distinguishable by the amplitude of signal produced by the label. The compounds are assembled from label components and protein cores to facilitate modular production of the compounds. In compounds containing two or more proteins, the proteins are typically covalently linked. Useful sets of compounds include sets of labeled nucleotide analogs, particularly dye-label nucleotide analogs that include tetravalent biotin-binding protein cores. 149-. (canceled)50. A composition comprising:a first compound comprising one or more tetravalent biotin-binding proteins and one or more label components bound to the one or more tetravalent biotin-binding proteins, each label component comprising one or more labels; anda second compound comprising two or more covalently linked tetravalent biotin-binding proteins and two or more label components bound to the two or more tetravalent biotin-binding proteins, each label component comprising one or more labels;wherein the total number of labels in the second compound is greater than that in the first compound.51. The composition of claim 50 , wherein the first compound comprises two or more covalently linked tetravalent biotin-binding proteins.52. The composition of claim 50 , wherein the label components on the first and second compounds are identical.53. The composition of claim 50 , wherein the labels are all identical.5455-. (canceled)56. The composition of claim 50 , wherein each label component is bound to the tetrameric biotin-binding protein through a biotin moiety.57. The composition of claim 50 , wherein each label component comprises a bis-biotin moiety bound to two biotin binding sites on one of the tetravalent biotin-binding proteins.58. The composition of claim 50 , wherein the one or more tetravalent biotin-binding proteins in the first compound and the ...

Recombinant polymerases with increased phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

MODIFIED BIOTIN-BINDING PROTEINS FOR IMMOBILIZATION

Compositions comprising covalently modified and mutated biotin-binding proteins, particularly biotin-binding proteins having a negative charge at physiological pH, are provided. Methods of producing such proteins are also provided, as are methods of immobilizing, sequencing, and making nucleic acids employing such proteins. 1. A composition comprising a modified biotin-binding protein that comprises one or more covalently attached sulfonate moieties.2. The composition of claim 1 , wherein the biotin-binding protein is a tetravalent biotin-binding protein.3. The composition of claim 1 , wherein the biotin-binding protein is streptavidin.4. The composition of claim 1 , wherein the biotin-binding protein comprises three or more covalently attached sulfonate moieties.5. The composition of claim 1 , wherein the biotin-binding protein comprises 12 or more covalently attached sulfonate moieties.67-. (canceled)8. The composition of claim 1 , wherein the biotin-binding protein comprises 45 or more covalently attached sulfonate moieties.9. (canceled)10. The composition of claim 1 , wherein the biotin-binding protein comprises one or more covalently attached 3 claim 1 ,4 claim 1 ,5-tris(3-sulfopropoxy)benzoyl moieties.1112-. (canceled)13. The composition of claim 1 , wherein the biotin-binding protein comprises 15 or more covalently attached 3 claim 1 ,4 claim 1 ,5-tris(3-sulfopropoxy)benzoyl moieties.14. (canceled)15. The composition of claim 1 , wherein the biotin-binding protein is a tetravalent biotin-binding protein comprising 45 or more covalently attached sulfonate moieties.16. The composition of claim 15 , wherein the biotin-binding protein is streptavidin.17. The composition of claim 1 , wherein the biotin-binding protein is a tetravalent biotin-binding protein comprising 15 or more covalently attached 3 claim 1 ,4 claim 1 ,5-tris(3-sulfopropoxy)benzoyl moieties.18. The composition of claim 17 , wherein the biotin-binding protein is streptavidin.19. The composition of ...

POLYMERASE ENZYME SUBSTRATES WITH PROTEIN SHIELD

Compositions and methods are provided for nucleotide analogs comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Nucleotide analogs of the invention have a protein shield between the dye moieties and nucleotide moieties of the analog. The protein prevents the direct interaction of the dye moiety with the enzyme carrying out nucleotide synthesis preventing photodamage to the enzyme. The nucleotide analogs of the invention can have multiple dyes and multiple nucleotide moieties. 126-. (canceled)27. A labeled nucleotide analog comprising:an avidin protein having four subunits, each subunit comprising one biotin binding site;one or two nucleotide components each comprising one or more phospholinked nucleotide moieties; andone or two dye components each comprising one or more dye moieties;wherein each component is bound to the avidin protein through a biotin moiety attached to a binding site on the avidin protein; andwherein at least one of the nucleotide or dye components comprise a bis-biotin moiety bound to two of the biotin binding sites on the avidin protein.28. The labeled nucleotide analog of wherein the nucleotide component comprises a bis-biotin moiety claim 27 , and the labeled nucleotide analog has two dye components each bound to the avidin through a single biotin moiety.29. The labeled nucleotide analog of wherein the dye component comprises a bis-biotin moiety claim 27 , and the labeled nucleotide analog has two nucleotide components each bound to the avidin through a single biotin moiety.30. The labeled nucleotide analog of wherein the labeled nucleotide analog has one dye component and one nucleotide component and each of the dye component and the nucleotide component comprises a bis-biotin moiety.31. The labeled nucleotide analog of wherein the number of dye moieties is between 1 and 18 and the number of nucleotide moieties is between 1 and 18.32. The labeled nucleotide analog of wherein the number of ...

RECOMBINANT POLYMERASES WITH INCREASED PHOTOTOLERANCE

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 120.-. (canceled)21. A composition comprising a φ29-type (phi29-type) recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:2 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position D17 , an amino acid substitution at position P305 , a Q96W substitution , a K128S substitution , an R258Q substitution , an K303Q substitution , a C452A substitution , an E463K substitution , an E463R substitution , and an E505Q substitution , wherein identification of positions is relative to SEQ ID NO:2 , and wherein said polymerase exhibits polymerase activity.22. The composition of claim 21 , wherein the recombinant polymerase comprises a P305L substitution claim 21 , wherein identification of positions is relative to SEQ ID NO:2.23. The composition of claim 21 , wherein the recombinant polymerase comprises one or more mutation or combination of mutations selected from the group consisting of an amino acid substitution at position 250 claim 21 , an amino acid substitution at position 372 claim 21 , an ...

Methods and Compositions for Enriching Compositions for Polymerase Enzyme Complexes

The present invention provides methods, compositions, and systems for enriching compositions for polymerase enzyme complexes. In particular, the methods, compositions, and systems of the present invention remove free polymerases from the compositions using one or more purification steps, including protease treatment, thus enriching the compositions for polymerases complexed with a template nucleic acid. 133-. (canceled)34. A method for enriching a loading composition for polymerase enzyme complexes , the method comprising:(a) providing a loading composition comprising free polymerases and polymerase enzyme complexes, wherein the polymerase enzyme complexes are in an inactive form;(b) applying a protease to the loading composition to remove free polymerases, thus enriching the loading composition for polymerase enzyme complexes; and(c) distributing the loading composition onto a substrate to provide a loaded substrate.35. The method of claim 34 , wherein the loading composition is purified to enhance the loading composition for polymerase enzyme complexes.36. The method of claim 34 , wherein applying step (b) is conducted after distributing step (c).37. The method of claim 34 , wherein the protease comprises an endopeptidase.38. The method of claim 34 , wherein the protease comprises trypsin.39. The method of claim 34 , wherein the substrate comprises an array of nanoscale wells.40. The method of claim 34 , wherein the distributing step (c) comprises immobilizing the polymerase enzyme complexes to the substrate.41. The method of claim 40 , wherein the polymerase enzyme complexes comprise a reactive element claim 40 , and the immobilizing occurs through an interaction of the reactive element and a binding site on the substrate.42. The method of claim 41 , wherein the reactive element comprises streptavidin and the binding site comprises biotin.43. The method of claim 41 , wherein the reactive element is resistant to cleavage by the protease.44. The method of claim 34 ...

Recombinant polymerases with increased phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Recombinant Polymerases with Increased Readlength and Stability for Single-Molecule Sequencing

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include increased stability, increased readlength, enhanced performance with large nucleotide analogs, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 1. A composition comprising a recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or at least 80% identical to SEQ ID NO:2 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position 429 , a C11A substitution , a Q99I substitution , an K337C substitution , a D570E substitution , a D570N substitution , a D570S substitution , a D570T substitution , a D570H substitution , a D570L substitution , a D570M substitution , a D570V substitution , a D570I substitution , a D570W substitution , a D570Y substitution , a D570F substitution , and a D570G substitution , wherein identification of positions is relative to SEQ ID NO:1.2. The composition of claim 1 , wherein the recombinant polymerase comprises an M429A substitution claim 1 , wherein identification of positions is relative to SEQ ID NO:1.3. The composition of claim 1 , wherein the recombinant polymerase ...

Laser Intensity Calibration

Systems, apparatuses, and methods are described for calibrating a laser power are described. A system may include a sample to be tested and a control sample that comprises a control analyte. A user may indicate a known concentration of the control analyte to the system (e.g., by entering a concentration value into a user interface or other process). The system may perform multiple runs at different laser powers and compare the measurements of each run against expected values for the control analyte at the known concentration. From that comparison, a calibrated laser power may be computed and that computed power level can be used by the system for the running of tests on an unknown sample. 1. A system comprising:a laser;a microfluidic channel;a sensor;a memory; and store a known concentration of a control analyte, the control analyte being present in a control sample at the known concentration;', 'determine, using the known concentration of the control analyte and an empirically determined model, at least one empirically determined characteristic of the control analyte;', 'perform multiple tests of the control sample at different laser powers applied to the microfluidic channel;', 'obtain measurements from the sensor while performing the multiple tests;', 'determine, based on the measurements from the sensor, at least one measured characteristic of the control analyte;', 'compute a calibrated laser power based on a comparison between the at least one measured characteristic and the at least one empirically determined characteristic; and', 'perform, at the calibrated laser power, at least one test of a sample., 'a processor configured to execute instructions stored in the memory, the instructions, when executed, cause the processor to2. The system according to claim 1 , wherein the sensor measurements are intensities of light generated by at least one of fluorescent dyes or fluorescent tags associated with components of the control sample in the microfluidic channel.3 ...

Heat exchange fluid purification for dialysis system

Disclosed herein are small, lightweight, portable, systems that have the capability of reliably, reproducibly, highly efficiently and relatively inexpensively providing a source of purified water of sufficient volumes for home dialysis. In addition, the systems disclosed herein require much less purified water at any one time than the volumes typically needed for dialysis today, thereby further reducing the expense of running the system at home.

DELAYING REAL-TIME SEQUENCING

Methods, compositions, and systems are provided that allow for reliable sequencing of the initial sequence region of a sequence of interest. The methods of the invention allow for more reliable barcoding of subpopulations of nucleic acids to be sequenced. 1. A method for delaying the sequencing of a sequence of interest in a single-molecule , real-time sequencing reaction comprising:simultaneously initiating a plurality of single-molecule, real-time sequencing reactions on a plurality of polymerase-template complexes, wherein each of the polymerase-template complexes comprises a polymerase enzyme and a nucleic acid template wherein the nucleic acid template comprises, in order, a priming region, a runway region, and an insert region comprising a sequence of interest, wherein the time from the initiation of the sequencing reactions to the time of sequencing the insert region is greater than about 30 seconds for a majority of the polymerase-template complexes.2. The method of wherein each priming region in each nucleic acid template in each of the polymerase-template complexes comprises a substantially identical sequence.3. The method of wherein each runway region in each nucleic acid template in each of the polymerase-template complexes comprises a substantially identical sequence.4. The method of wherein the template nucleic acid comprises a double-stranded region and a hairpin connecting the strands of the double-stranded region.5. The method of wherein the template nucleic acid comprises a double-stranded region and hairpins on each end of the double-stranded region connecting the two strands.6. The method of wherein the incorporation of nucleotide residues is detected using fluorescence.7. The method of wherein the incorporation of nucleotide residues is detected electronically or magnetically.8. The method of wherein the incorporation of nucleotide residues is detected by electrochemistry claim 1 , capacitance claim 1 , conductivity claim 1 , impedance claim 1 , ...

Laser enclosure

An enclosure for a laser in which on side of the enclosure includes a rotatable positioner and a vertically slidable door movable between open and closed positions. With the door in the open position, the positioner is free to rotate, thereby enabling an workstation to rotate into and out of the enclosure.

Wire feeder for welding

A wire conveying mechanism, preferably for a welding, cladding or additive manufacturing apparatus, with a slip clutch mechanism connectable to a motor. Such a wire conveying mechanism may include a wire feeder for feeding an electrode wire in a welding system having a drive roller assembly comprising a plurality of drive rollers to grip the electrode wire and to pull the electrode wire from an electrode wire source toward a conduit and a torque-managing device operatively coupled between a motor and the drive roller assembly, wherein the torque-managing device receives the rotational force from the first motor and regulates the first torque to output a second torque to the drive roller assembly.

MULTI-STAGE ACOUSTOPHORESIS DEVICE

An acoustophoresis device made up of modular components is disclosed. Several modules are disclosed herein, including ultrasonic transducer modules, input/output modules, collection well modules, and various connector modules. These permit different systems to be constructed that have appropriate fluid dynamics for separation of particles, such as biological cells, from a fluid. 1. A multi-stage acoustophoretic system , comprising: a flow chamber having at least one inlet and at least one outlet;', 'at least one ultrasonic transducer coupled to the flow chamber, the transducer including a piezoelectric material driven by a voltage signal to create a multi-dimensional acoustic standing wave in the flow chamber; and, 'three or more acoustophoretic devices fluidly connected to one another in series, each acoustophoretic device comprisinga reflector opposite from the at least one ultrasonic transducer.2. The multi-stage acoustophoretic system of claim 1 , wherein the three or more acoustophoretic devices are fluidly connected to one another by tubing.3. The multi-stage acoustophoretic system of claim 1 , comprising a total of four acoustophoretic devices.4. The multi-stage acoustophoretic system of claim 1 , further comprising a feed pump and a pump downstream of each acoustophoretic device.5. The multi-stage acoustophoretic system of claim 1 , further comprising a flowmeter upstream of the first acoustophoretic device and a flowmeter downstream of each acoustophoretic device.6. The multi-stage acoustophoretic system of claim 1 , wherein each acoustophoretic device has at least one dump diffuser acting as the inlet into the flow chamber.7. A method for continuously separating a second fluid or a particulate from a host fluid claim 1 , the method comprising: a flow chamber having at least one inlet and at least one outlet;', 'at least one ultrasonic transducer located on a wall of the flow chamber, the transducer including a piezoelectric material driven by a voltage ...

Recombinant polymerases for improved single molecule sequencing

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Methods and Compositions for Delivery of Molecules and Complexes to Reaction Sites

The present invention provides methods, compositions, and systems for distributing molecules and complexes into reaction sites. In particular, the methods, compositions, and systems of the present invention result in an active loading of molecules and complexes into reaction sites with improved efficiency over loading by passive diffusion methods alone. 177-. (canceled)79. The method of claim 78 , wherein the releasing step comprises applying a solution containing high salt to the coated surface.80. The method of claim 106 , wherein the releasing step comprises applying trypsin to the coated surface.81. The method of claim 78 , wherein the releasing step comprises applying a size exclusion resin to the coated surface.82. The method of claim 106 , wherein the positively charged peptide or polypeptide is a member selected from the group consisting of Tat claim 106 , poly-lysine claim 106 , poly-arginine claim 106 , and histone.83. The method of claim 106 , wherein the positively charged peptide or polypeptide comprises a Tat peptide.84. The method of claim 78 , wherein the nucleic acids are template nucleic acid molecules complexed with polymerase enzymes in polymerase enzyme complexes.85. The method of claim 84 , wherein the template nucleic acid molecules are further hybridized to a primer.86. The method of claim 84 , wherein the template nucleic acid molecules are at least 5 kb long.87. The method of claim 84 , wherein the template nucleic acid molecules are about 3 kb to about 20 kb in length.88. The method of claim 84 , wherein the delivering of the polymerase enzyme complexes is accomplished about 4 to about 100 times faster than seen with diffusion loading alone.89. The method of claim 84 , wherein the delivering of the polymerase enzyme complexes is accomplished at least 30 claim 84 , 40 claim 84 , 50 claim 84 , 60 claim 84 , 70 or 80 times faster than seen with diffusion loading alone.90. The method of claim 84 , wherein the delivering of the polymerase ...

Recombinant Polymerases For Improved Single Molecule Sequencing

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

METALWORKING SYSTEM WITH FORCE CONTROLLED WIRE FEED START OPERATION

A method for controlling a start of a metalworking operation. The method includes detecting an initial contact between a wire being fed from a welding apparatus and a workpiece and, in response to the detection, halting feeding of the wire from the welding apparatus. The method further includes activating a high energy heat source configured to heat a tip of the wire and resuming the feeding of the wire from the welding apparatus when the tip of the wire is heated by the high energy heat source to a plastic state. The feeding of the wire is resumed by measuring a force feedback from the wire contacting the workpiece. An apparatus for implementing the method is also disclosed.

SYSTEMS AND METHODS FOR ADAPTIVE CONTROL OF WIRE PREHEATING

A welding-type system includes a power supply configured to control preheating of an electrode wire. A controller is configured to receive a plurality of power values corresponding to a power output of the power supply and calculate an arc power value corresponding to an arc condition at the preheated electrode wire based on a rate of change of the plurality of power values. A target power output value is determined based on the calculated arc power value, and the power output is adjusted based on the determined target power value. 1. A welding-type system , comprising:a power supply configured to preheat an electrode wire; and receive a plurality of power values corresponding to a power output of the power supply;', 'calculate an arc power value corresponding to an arc condition at the preheated electrode wire based on a rate of change of the plurality of power values;', 'determine a target power output value based on the calculated arc power value; and', 'adjust the power output based on the determined target power value., 'a controller configured to2. The system as defined in claim 1 , wherein the power output is regulated between a high threshold value and a low threshold value about the target power output value.3. The system as defined in claim 2 , wherein the controller is configured to:compare a first power value of the plurality of power values to the high threshold power value and the low threshold power value; andadjust the power output based on the comparison.4. The system as defined in claim 2 , wherein the controller is configured to:raise the power output if the first power output is below the low threshold power value; andto lower the power output if the first power output is above the high threshold power value.5. The system as defined in claim 2 , further comprising a wire feeder claim 2 , the controller further configured to adjust a wire feed speed based on a comparison of the first power value to the target power output value and the high and ...

Recombinant Polymerases With Increased Phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 1. A composition comprising a recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or at least 80% identical to SEQ ID NO:2 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position 131 , an amino acid substitution at position 132 , a K135Q substitution , a K135S substitution , an H149D substitution , a Q183F substitution , a G197D substitution , a G197E substitution , an I201E substitution , a K206E substitution , an A437N substitution , and a D510S substitution , wherein identification of positions is relative to SEQ ID NO:1.2. The composition of claim 1 , wherein the recombinant polymerase comprises a K131E substitution or a K131Q substitution claim 1 , wherein identification of positions is relative to SEQ ID NO:1.3. The composition of claim 1 , wherein the recombinant polymerase comprises one or more mutation or combination of mutations selected from the group consisting of an amino acid substitution at position 253 claim 1 , an amino acid substitution at position 375 claim 1 , ...

Acoustic perfusion devices

Methods are disclosed for separating beads and cells from a host fluid. The method includes flowing a mixture containing the host fluid, the beads, and the cells through an acoustophoretic device having an ultrasonic transducer including a piezoelectric material driven by a drive signal to create a multi-dimensional acoustic standing wave. A drive signal is sent to drive the at least one ultrasonic transducer to create the multi-dimensional acoustic standing wave. A recirculating fluid stream having a tangential flow path is located substantially tangential to the standing wave and separated therefrom by an interface region. A portion of the cells pass through the standing wave, and the beads are held back from the standing wave in the recirculating fluid stream at the interface region. Also disclosed is an acoustophoretic device having a coolant inlet adapted to permit the ingress of a cooling fluid into the device for cooling the transducer.

FLUORESCENT POLYMERASE ENZYME SUBSTRATES HAVING PROTEIN SHIELDS

Compositions, methods, and systems are provided for fluorescent polymerase enzyme substrates comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Fluorescent polymerase enzyme substrates of the invention have a protein shield between the fluorescent dye moieties and nucleotide moieties of the polymerase enzyme substrate. The polymerase enzyme substrates have a nucleotide component and a dye component, each attached to a protein. The attachments can be covalent. The protein can, for example, prevent the direct interaction of the fluorescent dye moiety with the enzyme when carrying out nucleotide synthesis, preventing photodamage to the enzyme. The polymerase enzyme substrates of the invention can have multiple dyes and multiple nucleotide moieties. 1. A polymerase enzyme substrate comprising:a protein comprising at least 60 amino acids;a nucleotide component comprising at least one nucleoside polyphosphate attached through its phosphate portion to a first position on the protein;a dye component comprising at least one fluorescent dye moiety attached to a second position on the protein.2. The polymerase enzyme substrate of wherein the substrate is connected through covalent attachments.3. The polymerase enzyme substrate of wherein the protein comprises 60 to 1 claim 1 ,000 amino acids.4. The polymerase enzyme substrate of wherein the protein comprises 80 to 600 amino acids.5. The polymerase enzyme substrate of wherein the nucleotide component and dye component are covalently attached to the protein.6. The polymerase enzyme substrate of wherein the nucleotide component comprises two or more nucleoside phosphates.7. The polymerase enzyme substrate of wherein the substrate has 2 claim 1 , 3 claim 1 , 4 claim 1 , 5. 6 claim 1 , 7 claim 1 , or 8 nucleotide phosphates.8. The polymerase enzyme substrate of wherein the dye component comprises two or more fluorescent dye moieties.9. The polymerase enzyme substrate of wherein ...

Wide path welding, cladding, additive manufacturing

A welding or cladding apparatus in which one or more energy beam emitters are used to generate a wide beam spot transverse to a welding or cladding path, and one or more wide feeders feed wire to the spot to create a wide welding or cladding puddle.

LASER WELDING SYSTEMS FOR ALUMINUM ALLOYS AND METHODS OF LASER WELDING ALUMINUM ALLOYS

Systems and methods of a laser welding device to weld aluminum are disclosed. The device includes a laser generator to generate welding-type lasing power and a lens to focus the welding-type lasing power at a focal point on an aluminum workpiece to generate a weld puddle. A laser scanner to control the lens to move the focal point of the welding-type lasing power in multiple dimensions over the aluminum workpiece during welding, the laser generator and the laser scanner to perform the welding without filler metal being added to the workpiece.

FLUORESCENT POLYMERASE ENZYME SUBSTRATES HAVING PROTEIN SHIELDS

Compositions, methods, and systems are provided for fluorescent polymerase enzyme substrates comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Fluorescent polymerase enzyme substrates of the invention have a protein shield between the fluorescent dye moieties and nucleotide moieties of the polymerase enzyme substrate. The polymerase enzyme substrates have a nucleotide component and a dye component, each attached to a protein. The attachments can be covalent. The protein can, for example, prevent the direct interaction of the fluorescent dye moiety with the enzyme when carrying out nucleotide synthesis, preventing photodamage to the enzyme. The polymerase enzyme substrates of the invention can have multiple dyes and multiple nucleotide moieties. 125.-. (canceled)26. A method of making a labeled nucleotide analog comprising:(a) providing an avidin protein having four subunits, each subunit comprising one biotin binding site;(b) adding a nucleotide component, the nucleotide component comprising one or more phospholinked nucleotide moieties covalently connected to a bis-biotin moiety, whereby the bis-biotin moiety binds to two of the four subunits on the avidin protein, and(c) adding a dye component comprising one or more dye moieties covalently connected to a bis biotin moiety, whereby this bis-biotin moiety binds to the other two subunits on the avidin protein thereby forming a nucleotide analog.271. The method of claim wherein step c is performed after step b.281. The method of claim wherein step b is performed after step c.291. The method of claim wherein the number of dye moieties in the dye component is between 1 and 18.301. The method of claim wherein the number of nucleotide moieties in the nucleotide component is between 1 and 18.311. The method of claim wherein the number of dye moieties in the dye component is 1 , 2 , or 3 and the number of nucleotide moieties is 6 , 7 , or 8.321. The method of claim ...

Recombinant polymerases with increased phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Multi-amplitude modular labeled compounds

Sets of compounds bearing detectably different groups of labels are provided. Typically, different compounds bear different numbers of a single type of label and are thus distinguishable by the amplitude of signal produced by the label. The compounds are assembled from label components and protein cores to facilitate modular production of the compounds. In compounds containing two or more proteins, the proteins are typically covalently linked. Useful sets of compounds include sets of labeled nucleotide analogs, particularly dye-label nucleotide analogs that include tetravalent biotin-binding protein cores.

ACOUSTIC PERFUSION DEVICES

Acoustic perfusion devices for separating biological cells from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn. 1. An acoustic perfusion device , comprising:an acoustic chamber;an inlet port and an inlet flow path leading from the inlet port to the acoustic chamber;an outlet port and an outlet flow path leading from the acoustic chamber to the outlet port for recirculating a fluid mixture;at least one collection port for collecting harvest fluid from the fluid mixture; andat least one ultrasonic transducer and at least one reflector opposite the at least one ultrasonic transducer, wherein the at least one ultrasonic transducer includes a piezoelectric material driven by a voltage signal to create a multi-dimensional acoustic standing wave across a collection flow path leading from the acoustic chamber to the at least one collection port.2. The device of claim 1 , wherein the acoustic chamber is below the at least one collection port.3. The device of claim 1 , wherein the inlet flow path is shaped to generate a tangential flow path below an acoustic field generated by the acoustic standing wave.4. The device of claim 1 , wherein a pressure rise and an acoustic radiation force on cells are generated at an upstream interface region of the acoustic standing wave to clarify fluid passing through the acoustic standing wave.5. The device of claim 1 , wherein the at least one reflector is made of a transparent material.6. The device of claim 1 , wherein the outlet port is below the inlet port.7. The device of claim 1 , having a total of two or more ...

METHODS AND COMPOSITIONS FOR DELIVERY OF MOLECULES AND COMPLEXES TO REACTION SITES

The present invention provides methods, compositions, and systems for distributing molecules and complexes into reaction sites. In particular, the methods, compositions, and systems of the present invention result in loading of polymerase enzyme complexes into a predetermined number of reaction sites, including nanoscale wells. 1. A method for loading polymerase enzyme complexes into a predetermined number of nanoscale wells , the method comprising:(a) providing a surface comprising an array of nanoscale wells; (i) one or more nucleotides and/or nucleotide analogs; and', '(ii) polymerase enzyme complexes comprising a template nucleic acid and a polymerase enzyme,, '(b) contacting a loading solution to the surface, wherein the loading solution compriseswherein interactions between the nucleotides and/or nucleotide analogs and the polymerase enzyme complexes result in generation of signal pulses;(c) while the loading solution is in contact with the surface, monitoring the array of nanoscale wells to detect signal pulses from within the wells and thereby identifying nanoscale wells that have been loaded with a polymerase enzyme complex; and(d) maintaining the loading solution in contact with the surface until the predetermined number of nanoscale wells have been loaded with a polymerase enzyme complex.2. The method of claim 1 , wherein the loading solution comprises one or more labeled nucleotide analogs claim 1 , wherein the signal pulses are a result of non-incorporation events involving the labeled nucleotide analogs.3. The method of claim 2 , wherein the loading solution comprises one or more additives that maintain the polymerase enzymes in an inactive state.4. (canceled)5. The method of claim 3 , wherein the one or more additives comprise a divalent cation selected from the group consisting of strontium claim 3 , cobalt claim 3 , tin claim 3 , calcium claim 3 , nickel claim 3 , europium claim 3 , barium claim 3 , iron claim 3 , and zinc.6. The method of claim 2 , ...

Acoustic perfusion devices

Acoustic perfusion devices and processes for separating biomolecules from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn.

BIS-BIOTINYLATION TAGS

Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants. 1. A composition comprising a first reactant covalently coupled to a first bis-biotin tag , a second reactant covalently coupled to a second bis-biotin tag , and a streptavidin tetramer bound to both bis-biotin tags.2. The composition of claim 1 , wherein the first reactant is a protein.3. The composition of claim 2 , wherein the protein is a polymerase.4. The composition of claim 1 , wherein the first reactant is a label.5. The composition of claim 4 , wherein the label is a fluorescent label.6. The composition of claim 4 , wherein the label is a spin label.7. The composition of claim 1 , wherein the first reactant is a non-reactive component.8. The composition of claim 7 , wherein the label is a nucleic acid barcode.9. The composition of claim 7 , wherein the first reactant is an immobilization tag.10. The composition of claim 7 , wherein the first reactant is a purification tag.11. The composition of claim 1 , wherein the first reactant is a solid support.12. The composition of claim 11 , wherein the solid support is a bead.13. The composition of claim 11 , wherein the solid support is an array.14. The composition of claim 1 , wherein the first bis-biotin tag is tandemly oriented.15. The composition of claim 14 , wherein the first bis-biotin tag comprises two biotin moieties separated by 20-60 angstroms.16. The composition of claim 1 , wherein the streptavidin tetramer comprises a first streptavidin dimer bound to both biotin moieties of the first his-biotin tag claim 1 , and a second streptavidin dimer ...

Recombinant polymerases with increased phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Recombinant polymerases for incorporation of protein shield nucleotide analogs

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

COAXIAL LASER HOTWIRE HEAD

A laser processing head includes first and second contact points connected to a power source. The power source generates a current to flow through an electrode wire between the first and second contact points to heat the electrode wire. A laser source generates one or more laser beams having lasing power sufficient to at least partially melt the electrode wire. A coaxial laser head focuses the one or more laser beams at one or more focal points on a workpiece to at least partially melt the electrode wire. 1. A laser processing head , comprising:a first contact point connected to a power source;a second contact point connected to the power source, the power source configured to generate a current to flow through an electrode wire between the first contact point and the second contact point to heat the electrode wire;a laser source configured to generate one or more laser beams having lasing power sufficient to at least partially melt the electrode wire; anda coaxial laser head configured to focus the one or more laser beams at one or more focal points on a workpiece to at least partially melt the electrode wire.2. The laser processing head as defined in claim 1 , wherein the electrode wire is in electrical contact with both the first contact point and the second contact point.3. The laser processing head as defined in claim 1 , wherein the focal point corresponds to a location at which the electrode wire makes contact with the workpiece.4. The laser processing head as defined in claim 1 , wherein the power source comprises a controller configured to control a level of current flowing through the electrode wire between the first and second contact points to adjust a level of heating in the electrode wire.5. The laser processing head as defined in claim 4 , wherein the controller is further configured to receive a current measurement from a sensor in a current feedback loop and to control the level of current based on the current feedback loop.6. The laser processing ...

Modified biotin-binding proteins for immobilization

Compositions comprising covalently modified and mutated biotin-binding proteins, particularly biotin-binding proteins having a negative charge at physiological pH, are provided. Methods of producing such proteins are also provided, as are methods of immobilizing, sequencing, and making nucleic acids employing such proteins.

Recombinant polymerases with increased phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

LASER WELDING, CLADDING, AND/OR ADDITIVE MANUFACTURING SYSTEMS AND METHODS OF LASER WELDING, CLADDING, AND/OR ADDITIVE MANUFACTURING

Systems and methods of a laser welding device are disclosed. The laser welding device includes a laser generator configured to generate welding-type lasing power. A lens focuses the welding-type lasing power at a focal point on a workpiece to generate a puddle during a welding-type operation. A wire feeder is configured to feed wire to the puddle generated by the laser generator. A laser scanner controls the lens to move the focal point of the welding-type lasing power in multiple dimensions over the workpiece during the welding-type operation. In some examples, the feed wire is used in an additive manufacturing process. 1. A laser welding device , comprising:a laser generator configured to generate welding-type lasing power;a lens to focus the welding-type lasing power at a focal point on a workpiece to generate a puddle during a welding-type additive manufacturing operation;a wire feeder configured to feed wire to the puddle generated by the laser generator; anda laser scanner to control the lens to move the focal point of the welding-type lasing power in multiple dimensions over the workpiece during the welding-type additive manufacturing operation.2. The laser welding device as defined in claim 1 , wherein the laser scanner is configured to move the focal point in a circle claim 1 , an ellipse claim 1 , a zigzag claim 1 , a figure-8 claim 1 , a crescent claim 1 , a triangle claim 1 , a square claim 1 , a rectangle claim 1 , a non-linear pattern claim 1 , an asymmetrical pattern claim 1 , a pause claim 1 , or any combination thereof.3. The laser welding device as defined in claim 2 , wherein the movement of the focal point and relative movement between the workpiece and the laser scanner cause the lasing power to trace a superimposed pattern over the workpiece.4. The laser welding device as defined in claim 1 , wherein the laser scanner comprises a remote scan head with reflective optics or a rotary wedge scanner with transmissive optics.5. The laser welding ...

Recombinant polymerases for improved single molecule sequencing

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

RECOMBINANT POLYMERASES FOR INCORPORATION OF PROTEIN SHIELD NUCLEOTIDE ANALOGS

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Dispenser Control Systems and Methods

A method of operating a dispensing system having a material delivery cycle. In some embodiments, the material delivery cycle includes supplying water to a receptacle, performing an operation intended to release a material into the water, and delivering the material to a downstream component. The first step of the method is to initiate the material delivery cycle. Next, a conductivity proximate to the receptacle is monitored. Additionally, one or more error conditions are identified during the material delivery cycle based at least partially on the monitored conductivity. 1. A method of operating a dispensing system adapted to dispense material from a container , the method comprising:connecting the container to a receptacle at least partially contained within the dispensing system;supplying water to the receptacle;performing an operation intended to release material into the water from the container;monitoring a conductivity proximate the receptacle via a sensor positioned to sense conductivity in the receptacle;delivering the material and water from the receptacle through an outlet conduit of the dispensing system for use in cleaning operations by a downstream component;identifying one or more error conditions via a controller during at least one of the steps of supplying water to the receptacle, performing an operation intended to release material into the water, and delivering the material and water; anddetermining a location within the dispensing system where the one or more error conditions has occurred based on the identification of the one or more error conditions.2. The method of claim 1 , further comprising:supporting the container in the receptacle; anddelivering the material to a washing machine that is positioned downstream of the dispensing system.3. The method of claim 1 , wherein performing an operation intended to release material into the water comprises releasing a powder material or a granulated material into the water.4. The method of claim 1 , ...

RECOMBINANT POLYMERASES FOR IMPROVED SINGLE MOLECULE SEQUENCING

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 1. A composition comprising a recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or SEQ ID NO:2 , which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position A83 , an amino acid substitution at position C106 , an amino acid substitution at position A134 , an amino acid substitution at position R261 , an amino acid substitution at position R306 , an amino acid substitution at position W436 , and an amino acid substitution at position D476 , wherein identification of positions is relative to SEQ ID NO:1 , and which recombinant polymerase exhibits polymerase activity.2. The composition of claim 1 , wherein the recombinant polymerase comprises one or more mutation selected from the group consisting of a C106S substitution claim 1 , an A134S substitution claim 1 , an R261K substitution claim 1 , a W436Y substitution claim 1 , and a D476H substitution claim 1 , wherein identification of positions is relative to SEQ ID NO:1.3. The composition of claim 1 , wherein the recombinant polymerase comprises an amino acid sequence that is at least 90% ...

RECOMBINANT POLYMERASES WITH INCREASED PHOTOTOLERANCE

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 1. A composition comprising a φ29-type (phi29-type) recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:2 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position S191 , an amino acid substitution at position F436 , an amino acid substitution at position T438 , an amino acid substitution at position A444 , an amino acid substitution at position S524 , an L139R substitution , a D507Q substitution , and a T520H substitution , wherein identification of positions is relative to SEQ ID NO:2 , and wherein said polymerase exhibits polymerase activity.2. The composition of claim 1 , wherein the recombinant polymerase comprises one or more mutation selected from the group consisting of a T438I substitution claim 1 , a T438L substitution claim 1 , and an S524K substitution claim 1 , wherein identification of positions is relative to SEQ ID NO:2.3. The composition of claim 1 , wherein the recombinant polymerase comprises one or more mutation selected from the group consisting of an S191A ...

Delaying real-time sequencing

Methods, compositions, and systems are provided that allow for reliable sequencing of the initial sequence region of a sequence of interest. The methods of the invention allow for more reliable barcoding of subpopulations of nucleic acids to be sequenced.

Recombinant polymerases for incorporation of protein shield nucleotide analogs

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Recombinant polymerases with increased phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Recombinant polymerases with increased phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Recombinant Polymerases for Improved Single Molecule Sequencing

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 143-. (canceled)44. A composition comprising a recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or SEQ ID NO:2 , which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position V19 , an amino acid substitution at position C22 , an amino acid substitution at position L142 , an amino acid substitution at position Y224 , an amino acid substitution at position E239 , an amino acid substitution at position N409 , an E508R substitution , an E508K substitution , a D510K substitution , a D510R substitution , and an E515Q substitution , wherein identification of positions is relative to SEQ ID NO:1 , and which recombinant polymerase exhibits polymerase activity.45. The composition of claim 44 , wherein the recombinant polymerase comprises one or more mutation selected from the group consisting of a V19C substitution claim 44 , a C22V substitution claim 44 , an L142K substitution claim 44 , a Y224K substitution claim 44 , an E239G substitution claim 44 , and an N409C substitution claim 44 , wherein identification of positions is relative to SEQ ID NO:1.46 ...

LABELED NUCLEOTIDE ANALOGS HAVING PROTEIN SHIELDS

Compositions, methods, and systems are provided for nucleotide analogs comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Nucleotide analogs of the invention have a protein shield between the dye moieties and nucleotide moieties of the analog. The nucleotide analogs have two avidin proteins connected by a central dye component, and each avidin protein is further connected to a nucleotide component. The protein can prevent the direct interaction of the dye moiety with the enzyme carrying out nucleotide synthesis preventing photodamage to the enzyme. The nucleotide analogs of the invention can have multiple dyes and multiple nucleotide moieties.

Bis-biotinylation tags

Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants.

WIRE FEED LIMITER

A wire conveying mechanism, preferably for a welding, cladding or additive manufacturing apparatus, with a slip clutch mechanism connectable to a motor.

Recombinant polymerases with increased phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

ACOUSTIC PERFUSION DEVICES

Acoustic perfusion devices for separating biological cells from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn. 1. An acoustic cell retention device , comprising:a chamber;at least one ultrasonic transducer coupled to the chamber;at least one reflector opposite the at least one ultrasonic transducer across the chamber;the at least one ultrasonic transducer configured to be excited to generate a multi-dimensional acoustic standing wave with the at least one reflector, wherein the multi-dimensional acoustic standing wave is configured to retain cells and pass fluid in which the cells are entrained.2. The device of claim 1 , further comprising a recirculation path communicating with the acoustic chamber.3. The device of claim 1 , wherein the multi-dimensional acoustic standing wave is further configured to generate a pressure rise and an acoustic radiation force on the cells at an interface region of the multi-dimensional acoustic standing wave.4. The device of claim 3 , further comprising a locale adjacent to the interface region where the cell concentration is increased.5. The device of claim 1 , further comprising a locale in the multi-dimensional acoustic standing wave where the cell concentration is increased.6. The device of claim 1 , wherein the multi-dimensional acoustic standing wave is further configured to generate an acoustic radiation force with an axial force component and a lateral force component that are of the same order of magnitude.7. The device of claim 1 , further comprising a control system coupled to the at least one ultrasonic ...

Decorative illumination apparatus

A novel illumination apparatus that may be formed from any object of choice, such as a natural rock or the like. The apparatus is of simple construction and allows a user to easily adjust the amount of light to be emitted there from. Or the user may direct the light in a preferred direction, and optional accessories are also provided, such as the apparatus may include, a motion detector, speakers, a cup-holder, a secret compartment, etc.

ACOUSTIC PERFUSION DEVICES

Methods are disclosed for separating beads and cells from a host fluid. The method includes flowing a mixture containing the host fluid, the beads, and the cells through an acoustophoretic device having an ultrasonic transducer including a piezoelectric material driven by a drive signal to create a multi-dimensional acoustic standing wave. A drive signal is sent to drive the at least one ultrasonic transducer to create the multi-dimensional acoustic standing wave. A recirculating fluid stream having a tangential flow path is located substantially tangential to the standing wave and separated therefrom by an interface region. A portion of the cells pass through the standing wave, and the beads are held back from the standing wave in the recirculating fluid stream at the interface region. Also disclosed is an acoustophoretic device having a coolant inlet adapted to permit the ingress of a cooling fluid into the device for cooling the transducer. 1. A method for separating a desired target material from a host fluid , the method comprising: a flow chamber including at least one inlet and at least one outlet;', 'at least one ultrasonic transducer coupled to the flow chamber, the at least one ultrasonic transducer including a piezoelectric material configured to be driven to create a multi-dimensional acoustic standing wave in the flow chamber; and', 'a reflector located opposite from the at least one ultrasonic transducer; and, 'flowing a mixture containing the host fluid, beads, and the target material through an acoustophoretic device, the acoustophoretic device comprisingdriving the at least one ultrasonic transducer to create the multi-dimensional acoustic standing wave, wherein a recirculating fluid stream that includes a tangential flow path is located substantially tangential to the multi-dimensional acoustic standing wave and separated therefrom by an interface region;wherein the beads attach to the target material and are held back from the multi-dimensional ...

Laser welding, cladding, and/or additive manufacturing systems and methods of laser welding, cladding, and/or additive manufacturing

Systems and methods of a laser welding device are disclosed. The laser welding device includes a laser generator configured to generate welding-type lasing power. A lens focuses the welding-type lasing power at a focal point on a workpiece to generate a puddle during a welding-type operation. A wire feeder is configured to feed wire to the puddle generated by the laser generator. A laser scanner controls the lens to move the focal point of the welding-type lasing power in multiple dimensions over the workpiece during the welding-type operation. In some examples, the feed wire is used in an additive manufacturing process.

Methods and compositions for isolating template nucleic acids

The present invention is directed to methods and compositions for isolating template nucleic acids containing target sequences of interest, wherein those isolated template nucleic acids can be further assessed for information related to sequence and nucleic acid modifications.

METALWORKING SYSTEM WITH FORCE CONTROLLED WIRE FEED START OPERATION

A method for controlling a start of a metalworking operation. The method includes detecting an initial contact between a wire being fed from a welding apparatus and a workpiece and, in response to the detection, halting feeding of the wire from the welding apparatus. The method further includes activating a high energy heat source configured to heat a tip of the wire and resuming the feeding of the wire from the welding apparatus when the tip of the wire is heated by the high energy heat source to a plastic state. The feeding of the wire is resumed by measuring a force feedback from the wire contacting the workpiece. An apparatus for implementing the method is also disclosed. 127-. (canceled)28. A method for controlling a start of a metalworking operation comprising:detecting an initial contact between a workpiece and a wire fed from a wire feeding apparatus, wherein the detection comprises measuring a force on the wire; andin response to the detection, halting feeding of the wire from the welding apparatus.29. The method of claim 28 , further comprising:activating a high energy heat source while the wire is in contact with the workpiece, the high energy heat source configured to heat a tip of the wire to a softened state; andresuming the feeding of the wire from the wire feeding apparatus after activation of the high energy heat source.30. The method of claim 29 , further comprising detecting the softened state by measuring the force on the wire claim 29 , wherein the softened state is detected when the force is measured to be below a force threshold corresponding to the initial contact.31. The method of claim 29 , further comprising preheating the wire with a preheating heat source after the initial contact and prior to activating the high energy heat source.32. The method of claim 29 , wherein the high energy heat source is a laser or plasma.33. The method of claim 29 , further comprising maintaining a constant wire feed speed after resuming the feeding of the ...

Wire Feeder for Welding

A wire conveying mechanism, preferably for a welding, cladding or additive manufacturing apparatus, with a slip clutch mechanism connectable to a motor. Such a wire conveying mechanism may include a wire feeder for feeding an electrode wire in a welding system having a drive roller assembly comprising a plurality of drive rollers to grip the electrode wire and to pull the electrode wire from an electrode wire source toward a conduit and a torque-managing device operatively coupled between a motor and the drive roller assembly, wherein the torque-managing device receives the rotational force from the first motor and regulates the first torque to output a second torque to the drive roller assembly. 1. A wire feeder for feeding an electrode wire in a welding system , the wire feeder comprising:a drive roller assembly, the drive roller assembly comprising a plurality of drive rollers to grip the electrode wire and to pull the electrode wire from an electrode wire source and to push the electrode wire toward a conduit;a first motor to generate a rotational force having a first torque; anda torque-managing device operatively coupled between the first motor and the drive roller assembly,wherein the torque-managing device receives the rotational force from the first motor and regulates the first torque to output a second torque to the drive roller assembly.2. The wire feeder of claim 1 , wherein the drive roller assembly comprises a driving gear and a driven gear claim 1 , the torque-managing device being coupled to the driving gear.3. The wire feeder of claim 1 , wherein the second torque is greater than a feed torque and less than a bird nest torque.4. The wire feeder of claim 1 , wherein the torque-managing device is removably coupled with the first motor via a first drive shaft.5. The wire feeder of claim 4 , wherein the torque-managing device is removably coupled with the drive roller assembly via a second drive shaft.6. The wire feeder of claim 1 , wherein the torque- ...

METHODS, SYSTEMS, AND REAGENTS FOR DIRECT RNA SEQUENCING

Provided are compositions comprising recombinant polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for sequencing RNA or RNA/DNA templates. Polymerases that topologically encircle the template nucleic acid are provided. Also provided are methods of using such polymerases to make a DNA or to sequence a template comprising RNA. 1. A reaction mixture comprising a polymerase enzyme complex comprising a polymerase enzyme and a template nucleic acid that comprises RNA , wherein the polymerase topologically encircles the template nucleic acid.2. The reaction mixture of claim 1 , wherein the template nucleic acid is circular.3. The reaction mixture of claim 1 , wherein the template nucleic acid is an mRNA.4. The reaction mixture of claim 1 , comprising a primer hybridized to the template nucleic acid.5. The reaction mixture of claim 1 , wherein the polymerase enzyme complex is immobilized on a surface.6. The reaction mixture of claim 5 , comprising sequencing reagents in contact with the surface claim 5 , which sequencing reagents comprise reagents for carrying out nucleic acid synthesis including one or more labeled nucleotide analogs.7. The reaction mixture of claim 1 , comprising a phosphate-labeled nucleotide analog claim 1 , wherein the polymerase incorporates the nucleotide analog into a copy nucleic acid in response to the template nucleic acid.8. The reaction mixture of claim 1 , wherein the polymerase is a recombinant Φ29-type polymerase claim 1 , which recombinant polymerase comprises an amino acid sequence that is at least 70% identical to SEQ ID NO:1 or at least 70% identical to SEQ ID NO:2 claim 1 , and which recombinant polymerase comprises a cysteine residue at position V19 and a cysteine residue at position N409 claim 1 , wherein identification of positions is relative to SEQ ID NO:1 claim 1 , wherein the side chains of the cysteine residues at positions V19 ...

HEAT EXCHANGE FLUID PURIFICATION FOR DIALYSIS SYSTEM

Disclosed herein are small, lightweight, portable, systems that have the capability of reliably, reproducibly, highly efficiently and relatively inexpensively providing a source of purified water of sufficient volumes for home dialysis. In addition, the systems disclosed herein require much less purified water at any one time than the volumes typically needed for dialysis today, thereby further reducing the expense of running the system at home. 1. A water treatment device , comprising:at least first and second laminae arranged in a stacked relationship so as to form a stack of laminae, each lamina having a fluid flow field, the fluid flow field of the first lamina configured for receiving blood and the fluid flow field of the second lamina configured for receiving dialysate;a first cut-out in the first lamina, wherein the first cut-out surrounds at least a portion of the fluid flow field of the first lamina;a second cut-out on the second lamina, wherein the second cut-out surrounds at least a portion of the fluid flow field of the second lamina, and wherein the first and second cut-outs vertically align with one another in the stack to form a collective cut-out that extends through the stack; anda first plate on the top of the stack and a second plate on the bottom of the stack, wherein the first and second plates enclose the top and bottom of the collective cut-out creating an insulating chamber to the fluid flow fields of the stack.2. The device of claim 1 , wherein each cut-out surrounds a respective flow field along at least three sides of the flow field.3. The device of claim 1 , wherein each cut-out surrounds at least eighty percent of the respective flow field.4. The device of claim 1 , wherein the stack includes more than two laminae.5. The device of claim 1 , wherein the collective cut-out forms a U-shape around the fluid flow fields of the stack.6. The device of claim 1 , further comprising at least one heat transfer layer interleaved between the first ...

Methods and Compositions for Isolating Template Nucleic Acids

The present invention is directed to methods and compositions for isolating template nucleic acids containing target sequences of interest, wherein those isolated template nucleic acids can be further assessed for information related to sequence and nucleic acid modifications. 1. A method for isolating template nucleic acids containing one or more target sequences , said method comprising:(a) providing a population of circular template nucleic acids, wherein a plurality of said population of circular template nucleic acids comprises said one or more target sequences; (i) one of said population of circular template nucleic acids, and', '(ii) a linear nascent strand comprising at least one copy of a sequence complementary to said one or more target sequences;, '(b) amplifying said population of template nucleic acids with a strand displacing polymerase to produce amplified template nucleic acids, wherein said amplified template nucleic acids each comprise(c) applying one or more affinity handles to said amplified template nucleic acids, wherein said one or more affinity handles associate with said at least one copy of a sequence complementary to said one or more target sequences;(d) separating amplified template nucleic acids associated with said affinity handles from amplified template nucleic acids not associated with said affinity handles;thereby isolating said template nucleic acids containing one or more target sequences.2. The method of claim 1 , wherein said population of circular template nucleic acids comprises genomic DNA.3. The method of claim 1 , wherein said circular template nucleic acids comprise a single-stranded portion and a double-stranded portion.4. The method of claim 1 , wherein said circular template nucleic acids are single stranded or double stranded.5. The method of claim 1 , wherein said template nucleic acids comprise:(a) a first strand segment;(b) a second strand segment substantially complementary to said first strand segment;(c) a first ...

Managing a Fleet of Electronic Devices

This disclosure describes, in part, techniques for managing access to a fleet account of mobile electronic devices. For instance, an administrator of a business and educational institution may place an order with an offering service to purchase a large number of eBook reader devices. The offering service identifies that the business and educational institution is ordering a fleet of devices. Furthermore, each of the devices ordered are associated or registered to a managed or fleet account controlled by the administrator of the business and educational institution. The techniques described herein prevent an individual user of a fleet device from de-registering the device from the managed account, and also allow an account administrator to alter preferred settings of the managed account. Further, techniques are described for allowing a user of a device to opt-in or associate their device with the account. 1. A method comprising: receiving an order for one or more client devices;', 'determining whether the one or more client devices are to be associated with an account;', associating each of the one or more client devices with the account; and', 'sending configuration instructions to each of the one or more client devices, the configuration instructions adapted to modify a setting of each of the one or more client devices to prevent the respective client device from disassociation from the account,', 'wherein the account is associated with specified content, the specified content being accessible by each client device that is associated with the account., 'at least partly in response to determining that the one or more client devices are to be associated with the account], 'under control of one or more computing systems configured with executable instructions,'}2. A method as recited in claim 1 , wherein the account is associated with specified content claim 1 , the specified content being accessible by each client device that is associated with the account.3. A ...

Context Aware Content Distribution

Described herein are systems and methods for context aware content distribution. Content such as e-books, presentations, audio, video, applications, and so forth may be distributed to user devices after establishing a trust relationship. An invitation to provide content may be sent to a user device based on the context of the user device, such as proximity to pre-determined devices, location, and so forth. Upon acceptance of the invitation to opt-in to receiving content, a trust relationship may be established which provides for the transfer of content to the user device. 1. A computer-implemented method for utilizing processing resources of a computerized system , the computer-implemented method comprising:receiving context data from a trusted user device, wherein the context data includes information indicative of a location of the trusted user device;identifying, based at least in part on the information indicative of the location, an invitation associated with a trusted content provider, wherein the invitation includes information indicative of content available from the trusted content provider to the trusted user device at the location and information indicative to provide access to the content;sending the invitation to the trusted user device;receiving an indication of acceptance of the invitation from the trusted user device;based at least in part on the acceptance, establishing a trust relationship between the trusted user device and the trusted content provider; andbased at least in part on the trust relationship, generating instructions configured to initiate transfer of content from the trusted content provider to the trusted user device.2. The method of claim 1 , the location comprising one or more of a geographic location claim 1 , a relative location based at least in part on a description associated with the relative location claim 1 , or a geographic or relative location at which one or more pre-determined devices are present.3. The method of claim ...

WELDING SYSTEM WITH ARC CONTROL

A method and apparatus for controlling arc/short between a wire and a work piece is described. A current path parallel to the wire/work is provided. The voltage drop across the parallel path can be preset, to limit the wire/work voltage, or it can be controlled to a desired level. The control can be in response to feedback. 1. A welding-type system , comprising:a power circuit disposed to provide welding-type power on a first electrode and a second electrode, wherein the power circuit has at least one control input;an arc clamp module, electrically coupled to the first electrode and the second electrode; anda controller, having a power circuit control output connected to the at least one control input.2. The welding-type system of claim 1 , wherein the arc clamp module is an active arc clamp module and has an arc clamp control input claim 1 , and wherein the controller has an arc clamp control module having an arc clamp control output connected to the arc clamp control input.3. The welding-type system of claim 2 , further comprising a feedback circuit electrically coupled to at least one of the first electrode claim 2 , second electrode claim 2 , and arc clamp module claim 2 , wherein the feedback circuit is disposed to provide a feedback signal to the controller claim 2 , wherein the arc clamp control output is responsive to the feedback signal.4. The welding-type system of claim 3 , wherein the feedback circuit includes a current sensor claim 3 , and wherein the arc clamp control output is responsive to the current sensor.5. The welding-type system of claim 4 , wherein the feedback circuit includes a voltage sensor claim 4 , and wherein the arc clamp control output is responsive to the voltage sensor.6. The welding-type system of claim 3 , wherein the feedback circuit includes a voltage sensor claim 3 , and wherein the arc clamp control output is responsive to the voltage sensor.7. The welding-type system of claim 2 , wherein the arc clamp control module is at ...

Synergic air compressor for gouging in a multi-function machine

A multi-function machine is provided. In one embodiment, the multi-function machine includes a generator driven by an engine for generating electrical power. The multi-function machine also includes a welding power supply electrically powered by the generator to provide an arc gouging output during an arc gouging process when the multi-function machine is in an arc gouging mode. The multi-function machine further includes an air compressor system driven by the generator to provide a compressed air output to support the arc gouging process. The multi-function machine also includes a controller operatively connected to the welding power supply and the air compressor system. The air compressor system is commanded by the controller to activate to provide the compressed air output when the arc gouging mode is selected and/or change a characteristic of the compressed air output when an arc gouging set point of the arc gouging mode is changed.

WELDING SYSTEM WITH ARC CONTROL

A method and apparatus for controlling arc/short between a wire and a work piece is described. A current path parallel to the wire/work is provided. The voltage drop across the parallel path can be preset, to limit the wire/work voltage, or it can be controlled to a desired level. The control can be in response to feedback. 1. A welding-type system , comprising:a power circuit disposed to provide welding-type power on a first electrode and a second electrode, wherein the power circuit has at least one control input;an arc clamp module, electrically coupled to the first electrode and the second electrode; anda controller, having a power circuit control output connected to the at least one control input.2. The welding-type system of claim 1 , wherein the arc clamp module is an active arc clamp module and has an arc clamp control input claim 1 , and wherein the controller has an arc clamp control module having an arc clamp control output connected to the arc clamp control input.3. The welding-type system of claim 2 , further comprising a feedback circuit electrically coupled to at least one of the first electrode claim 2 , second electrode claim 2 , and arc clamp module claim 2 , wherein the feedback circuit is disposed to provide a feedback signal to the controller claim 2 , wherein the arc clamp control output is responsive to the feedback signal.4. The welding-type system of claim 3 , wherein the feedback circuit includes a current sensor claim 3 , and wherein the arc clamp control output is responsive to the current sensor.5. The welding-type system of claim 4 , wherein the feedback circuit includes a voltage sensor claim 4 , and wherein the arc clamp control output is responsive to the voltage sensor.6. The welding-type system of claim 3 , wherein the feedback circuit includes a voltage sensor claim 3 , and wherein the arc clamp control output is responsive to the voltage sensor.7. The welding-type system of claim 2 , wherein the arc clamp control module is at ...

Scene flow estimation using shared features

Scene flow represents the three-dimensional (3D) structure and movement of objects in a video sequence in three dimensions from frame-to-frame and is used to track objects and estimate speeds for autonomous driving applications. Scene flow is recovered by a neural network system from a video sequence captured from at least two viewpoints (e.g., cameras), such as a left-eye and right-eye of a viewer. An encoder portion of the system extracts features from frames of the video sequence. The features are input to a first decoder to predict optical flow and a second decoder to predict disparity. The optical flow represents pixel movement in (x,y) and the disparity represents pixel movement in z (depth). When combined, the optical flow and disparity represent the scene flow.

WIDE PATH WELDING, CLADDING, ADDITIVE MANUFACTURING

A welding or cladding apparatus in which one or more energy beam emitters are used to generate a wide beam spot transverse to a welding or cladding path, and one or more wide feeders feed wire to the spot to create a wide welding or cladding puddle. 1. A metalworking apparatus , comprising:one or more energy beam emitters arranged to irradiate a workpiece and have one or more spots of high energy incident on the workpiece; andone or more wire feeders configured to feed one or more wires to the one or more spots and which when melted by the energy beam or beams form a welding puddle, each spot having a width in a direction transverse to a direction of a welding path, the cumulative width of the one or more spots being greater than a length of the one or more welding spots in the direction of the welding path.2. The apparatus of claim 1 , comprising a plurality spots continuously arrayed along a straight line.3. The apparatus of claim 2 , wherein the width direction of the one or more spots is orthogonal to the welding path.4. The apparatus of claim 2 , wherein the width direction of the one or more spots is at an oblique angle to the welding path.5. The apparatus of claim 1 , wherein the cumulative width of the spots is 4 mm or greater.6. The apparatus of claim 1 , comprising a single energy beam emitter which emits a wide energy beam or a moving energy beam to create a wide welding puddle.7. The apparatus of claim 1 , comprising a plurality energy beam emitters which emit energy beams of substantially equal cross-section and intensity.8. The apparatus of claim 1 , comprising a plurality energy beam emitters which emit energy beams which different energy intensities.9. The apparatus of claim 1 , wherein each energy beam emitter is a laser.10. The apparatus of claim 5 , wherein an intensity of the energy varies across the spot.11. A metalworking process comprising:irradiating a workpiece with one or more beams of energy and creating one or more spots of high energy ...

SEAL STRUCTURE AND METHOD OF FORMING

A seal structure can include a protruding portion of an outer surface of a body. An inner surface of the protruding portion may define at least a portion of a channel. The protruding portion may be configured to engage and at least partially deform into the channel in response to a load generated by pressing an outer surface of the protruding portion against a contact surface of a contact member to provide a seal between the body and the contact member. The seal structure may be integrally formed within the body as a monolithic object. The body may face towards the contact surface of the contact member during operation of the seal structure. A method may include forming a monolithic body with an integral seal structure by additively fusing material so that the body has an outer surface facing towards a contact surface of a contact member. 1. A seal structure , comprising:a protruding portion of an outer surface of a body, with an inner surface of the protruding portion defining at least a portion of a channel, the protruding portion being configured to engage and at least partially deform into the channel in response to a load generated by pressing an outer surface of the protruding portion against a contact surface of a contact member to provide a seal between the body and the contact member; andthe seal structure is integrally formed within the body as a monolithic object, and the body is configured to face towards the contact surface of the contact member during operation of the seal structure.2. The seal structure of claim 1 , wherein the protruding portion is resiliently biased against the load when in operation.3. The seal structure of claim 2 , wherein the channel is at least partially hollow and contains a biasing material.4. The seal structure of claim 3 , wherein the biasing material comprises at least one of: a series of support structures that extend at least part of the way from a floor of the channel to the inner surface claim 3 , and a compressible ...

ACCOUSTIC MASKING SYSTEM AND METHOD FOR ENABLING HIPAA COMPLIANCE IN TREATMENT SETTING

A method and system for masking to a listener a conversation between a care provider and a patient such that the conversation becomes unintelligible to the listener, includes interposing an acoustically absorbent curtain that substantially spans between the floor and the ceiling and is positioned between the listener and the care provider and patient. A masking device is affixed to the curtain generally between the floor and the ceiling at a position along the barrier length and at a height approximately coinciding with a height of a mouth of the care provider. The masking device includes at least one speaker having an axis oriented generally perpendicular to the plane the curtain defines. An amplifier drives the speaker to produce a sound to propagate along the axis. A signal source provided at the amplifier input produces the sound selected to mask the conversation between the care provider and the patient. 1. A method for masking , to a listener , a conversation between a care provider and a patient such that the conversation becomes unintelligible to the listener , the conversation occurring within a space defined by a floor and a ceiling , the method comprising:interposing an acoustically absorbent curtain that substantially spans between the floor and the ceiling and is positioned between a treatment group, including the care provider and the patient, and the listener, the acoustically absorbent curtain defining a barrier length; at least one speaker having an axis oriented generally perpendicular to the plane the curtain defines; and', 'an amplifier to drive the speaker to produce a sound to propagate along the axis, the sound being defined according to a signal at an amplifier input; and, 'providing a masking device affixed to the curtain generally between the floor and the ceiling and at a height approximately coinciding with a height of a mouth of the care provider, the masking device includingproviding a masking signal from a signal source to the ...

METHODS AND COMPOSITIONS FOR LOADING OF POLYMERASE COMPLEXES

The present disclosure provides methods, compositions, and systems for distributing polymerase compositions into array regions. In particular, the described methods, compositions, and systems utilize density differentials and/or additives to increase efficiency in the distribution of polymerase compositions to a surface as compared to methods utilizing only diffusion control. 2. The method of claim 1 , wherein the spike solution comprises a neutral and hydrophilic polysaccharide.3. The method of claim 1 , wherein the spike solution comprises a highly branched claim 1 , high-mass polysaccharide.4. The method of claim 1 , wherein the spike solution comprises a volume excluding buffer.5. The method of claim 1 , wherein the spike solution comprises an additive selected from the group consisting of dextran claim 1 , aminodextran claim 1 , dextrin claim 1 , cluster dextrin claim 1 , Ficoll claim 1 , polyetheylene glycol claim 1 , sucrose claim 1 , DMSO claim 1 , glycerol claim 1 , and pullulan.6. The method of claim 1 , wherein the spike solution comprises Ficoll.7. The method of claim 1 , wherein the distributing occurs at about a 2 to about a 25-fold faster rate as compared to distributing without the spike solution.8. The method of claim 1 , wherein the distributing occurs at about a 5 to about a 20-fold faster rate as compared to distributing without the spike solution.9. The method of claim 1 , wherein the distributing occurs at about a 10 to about a 15-fold faster rate as compared to distributing without the spike solution.10. The method of claim 1 , wherein the distributing occurs at least 5 times faster as compared to distributing without the spike solution.11. The method of claim 1 , wherein the distributing occurs at least 10 times faster as compared to distributing without the spike solution having a higher density than the buffer.12. The method of claim 1 , wherein the distributing occurs at least 15 times faster as compared to distributing without the spike ...

LASER ENCLOSURE

An enclosure for a laser in which on side of the enclosure includes a rotatable positioner and a vertically slidable door movable between open and closed positions. With the door in the open position, the positioner is free to rotate, thereby enabling an workstation to rotate into and out of the enclosure. 1. An enclosure with one side comprising:at least one vertically sliding door movable between open and closed positions and having a first edge;a positioner rotatable around a vertical axis; anda wall member carried on the positioner, the wall member also having a second edge, the wall member is rotatable with the positioner from parallel to a plane of the vertically sliding door to out of the plane,', 'the edges of the vertically sliding door and the wall member having mating profiles., 'wherein,'}2. The enclosure of claim 1 , wherein the first edge or the second edge includes an L-shaped or a U-shaped channel into which the second edge or first edge is received claim 1 , respectively.3. The enclosure of claim 1 , further comprising at least one flange extending along a plane of either the wall member or the vertically sliding door and from an edge of the wall member or the vertically sliding door claim 1 , respectively claim 1 , so as to at least define an L-shaped channel claim 1 , 'when the vertically sliding door member is in the closed position, the edge of the vertically sliding door or the edge of the wall member, respectively, is received within the channel and the flange overlaps a gap between the edges of the vertically sliding door member and the wall member.', 'wherein,'}4. The enclosure of claim 3 , wherein the flange extends along a plane of the wall member and from an edge of the wall member.5. The enclosure of claim 3 , wherein the flange extends along a plane of the vertically sliding door and from an edge of the vertically sliding door.6. The enclosure of claim 1 , comprising two flanges claim 1 , each extending from an edge and along respective ...

TECHNIQUES FOR TRANSFERRING DATA ACROSS AIR GAPS

Techniques for transferring data to a secure computing region that is isolated from any public networks is disclosed. In some embodiments, one or more artifacts for the secure computing region are packaged. As part of the packaging, one or more data packets and metadata for the one or more artifacts can be generated. The metadata indicates corresponding destination components for the one or more artifacts within the secure computing region. The data packet(s) are received and transmitted by a first data diode endpoint device. The first data diode endpoint device is configured to transmit data to the secure computing region via a second data diode endpoint device, the second data diode endpoint device being configured to restrict data from being transmitted to a destination outside of the secure computing region. The transmission of the one or more data packets may be monitored by the system. 1. A computer-implemented method , comprising:packaging, by a component of a computing system managed by a declarative infrastructure provisioner, one or more artifacts for a secure computing region that is isolated from any public networks, the packaging comprising generating one or more data packets and metadata for the one or more artifacts, the metadata indicating one or more destination components for the one or more artifacts within the secure computing region;receiving, by a first data diode endpoint device of a computing system, the one or more data packets comprising the one or more artifacts and metadata, the first data diode endpoint device being configured to transmit package data to the secure computing region via a second data diode endpoint device, the second data diode endpoint device being configured to restrict data from being transmitted to a destination outside of the secure computing region;transmitting, by the first data diode endpoint device, the one or more data packets to the second data diode endpoint device; andmonitoring, by the first data diode ...

Recombinant Polymerases With Increased Phototolerance

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 124.-. (canceled)25. A composition comprising a φ29-type recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position E20 , an amino acid substitution at position R308 , a Q99W substitution , a K131S substitution , an R261Q substitution , a C290F substitution , an R306Q substitution , a C455A substitution , an E466K substitution , an E466R substitution , and an E508Q substitution , wherein identification of positions is relative to SEQ ID NO:1 , and wherein said polymerase exhibits polymerase activity.26. The composition of claim 25 , wherein the recombinant polymerase comprises an E20D substitution or an R308L substitution claim 25 , wherein identification of positions is relative to SEQ ID NO:1.27253375484512510148224239250437235515141142504508513523536539205472437253508510. The composition of claim 25 , wherein the recombinant polymerase comprises one or more mutation or combination of mutations selected from the group consisting of ...

RECOMBINANT POLYMERASES FOR INCORPORATION OF PROTEIN SHIELD NUCLEOTIDE ANALOGS

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 127-. (canceled)28. A composition comprising: a recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO: 1 or at least 80% identical to SEQ ID NO:2 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position T434 , an amino acid substitution at position A435 , an amino acid substitution at position I452 , an A83E substitution , an A83R substitution , and a K539E substitution , wherein identification of positions is relative to SEQ ID NO: 1.29. The composition of claim 28 , wherein the recombinant polymerase comprises one or more mutation selected from the group consisting of a T434A substitution claim 28 , a T434S substitution claim 28 , an A435S substitution claim 28 , and an I452F substitution claim 28 , wherein identification of positions is relative to SEQ ID NO:1.30. The composition of claim 28 , wherein the recombinant polymerase comprises one or ...

RECOMBINANT POLYMERASES WITH INCREASED PHOTOTOLERANCE

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 124.-. (canceled)25. A composition comprising a Φ29-type (phi29-type) recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or at least 80% identical to SEQ ID NO:2 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position S194 , an amino acid substitution at position Y439 , an amino acid substitution at position T441 , an amino acid substitution at position A447 , an amino acid substitution at position S527 , an amino acid substitution at position K539 , an L142R substitution , a D510Q substitution , and a D523H substitution , wherein identification of positions is relative to SEQ ID NO:1 , and wherein said polymerase exhibits polymerase activity.26. The composition of claim 25 , wherein the recombinant polymerase comprises one or more mutation selected from the group consisting of a T441I substitution claim 25 , a T441L substitution claim 25 , an S527K substitution claim 25 , and a K539Q substitution claim 25 , wherein identification of positions is relative to SEQ ID NO:1.27. The ...

RECOMBINANT POLYMERASES FOR IMPROVED SINGLE MOLECULE SEQUENCING

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 1. A composition comprising a recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or SEQ ID NO:2 , which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position A68 , an amino acid substitution at position A83 , an amino acid substitution at position C106 , an amino acid substitution at position A134 , an amino acid substitution at position R261 , an amino acid substitution at position R306 , an amino acid substitution at position W436 , an amino acid substitution at position D476 , and an amino acid substitution at position P558 , wherein identification of positions is relative to SEQ ID NO: 1 , and which recombinant polymerase exhibits polymerase activity.2. The composition of claim 1 , wherein the recombinant polymerase comprises one or more mutation selected from the group consisting of an A68S substitution claim 1 , a C106S substitution claim 1 , an A134S substitution claim 1 , an R261K substitution claim 1 , a W436Y substitution claim 1 , a D476H substitution claim 1 , and a P558A substitution claim 1 , wherein identification of positions ...

Systems and methods for hybrid laser and arc welding additive manufacturing

Disclosed is a hybrid additive manufacturing system that includes a laser system and an additive manufacturing tool, such as an arc welding type torch. The tool is configured to receive a metallic electrode wire, which is heated by a power supply to create droplets for deposition to create the part by building up successive layers of metal. The additive manufacturing system operates through coordination of the laser system to generate a laser beam, which is applied to a weld bead, and an arc welding process, which provides material for the part. A threshold value of laser intensity and/or power can be applied to the weld puddle to stabilize the arc. Through the laser beam, an arc cone position can be manipulated such that the energy into the molten pool can be redistributed.

ACOUSTIC PERFUSION DEVICES

Acoustic perfusion devices and processes for separating biomolecules from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn. 1. A process for separating biomolecules from a fluid mixture , the process comprising: an acoustic chamber that receives the fluid mixture containing the biomolecules and cells; and', 'an ultrasonic transducer that includes a piezoelectric material that can be excited to generate a multi-dimensional acoustic standing wave in the acoustic chamber;, 'flowing a fluid mixture containing biomolecules and cells through an acoustic perfusion device, the acoustic perfusion device comprisingexciting the ultrasonic transducer to create the multi-dimensional acoustic standing wave; andretaining cells with a larger size in the fluid mixture with the multi-dimensional acoustic standing wave.2. The process of claim 1 , wherein the cells with a larger size are trapped by the multi-dimensional acoustic standing wave and the biomolecules pass through the multi-dimensional acoustic standing wave.3. The process of claim 2 , wherein the trapped cells are recycled to a bioreactor upstream of the acoustic perfusion device.4. The process of claim 1 , wherein the biomolecules are therapeutic antibodies.5. The process of claim 4 , wherein the fucosylation of the therapeutic antibodies is increased and the efficacy of the therapeutic antibodies is changed.6. The process of claim 1 , wherein the biomolecules are produced by culturing cells in a bioreactor prior to flowing the fluid mixture through the acoustic perfusion device.7. The process of claim 1 , ...

ACOUSTIC PRE-CONDITIONER

Devices and methods for pre-conditioning and/or post-conditioning a host fluid containing a second fluid or particulate are disclosed. The devices include a flow chamber having first opening and a particulate outlet. The devices can also include side openings and alignment, fluid, and particulate screens. An ultrasonic transducer can be driven to create an acoustic standing wave in the flow chamber, or alternatively be driven to excite the wall of the flow chamber in which it is located. This creates a uniformly stratified flow within the flow chamber, with the second fluid or particulate being aligned in planes in the fluid mixture. This permits the host fluid to be separated therefrom using the fluid screen and the particulate screen. 1. An acoustophoretic device , comprising:a flow chamber having a particulate outlet at a first end of the flow chamber and a first opening at a second end of the flow chamber opposite the first end thereof;at least one ultrasonic transducer located on a wall of the flow chamber, the at least one ultrasonic transducer including a piezoelectric material driven by a voltage signal to create an acoustic standing wave in the flow chamber;a reflector located on a wall on the opposite side of the flow chamber from the at least one ultrasonic transducer;at least one side opening located on a wall of the flow chamber between the reflector and the at least one ultrasonic transducer;a fluid screen located between the particulate outlet and the flow chamber, the fluid screen including a plurality of slots therein.2. The acoustophoretic device of claim 1 , wherein the slots in the outlet screen have a width equal to about one-quarter of the wavelength of the acoustic standing wave.3. The acoustophoretic device of claim 1 , wherein the slots in the outlet screen have a width of between about 0.005 inches and 0.02 inches and a height of between about 0.25 inches and 0.75 inches.4. The acoustophoretic device of claim 1 , further comprising at least ...

PREDICTIVE MAINTENANCE

Methods, computer program products, and systems are provided herein for providing predictive maintenance for an industrial cleaning device. An example method includes receiving operational data indicating operational parameters associated with the industrial cleaning device during operation thereof. The method further includes identifying a trend in the operational data. The method further includes determining that a particular fault event associated with the industrial cleaning device is likely to occur within a particular time frame. The method further includes providing an alert indicating that the particular fault event is likely to occur within the particular time frame. 1. A method for providing predictive maintenance for an industrial cleaning device , the method comprising:receiving operational data indicating operational parameters associated with the industrial cleaning device during operation thereof;identifying a trend in the operational data;determining that a particular fault event associated with the industrial cleaning device is likely to occur within a particular time frame; andproviding an alert indicating that the particular fault event is likely to occur within the particular time frame.2. The method of further comprising claim 1 , in response to the determining claim 1 , ordering at least one of a part or a supply for the industrial cleaning device wherein the ordered part or supply is selected based on the particular fault event.3. (canceled)4. The method of further comprising claim 1 , in response to the determining claim 1 , scheduling maintenance for the industrial cleaning device to prevent the fault event claim 1 , wherein the maintenance is scheduled to occur prior to the end of the particular time frame.5. (canceled)6. The method of further comprising claim 1 , in response to the determining claim 1 , providing information relating to a short term solution for the particular fault event.7. The method of claim 6 , wherein the short term ...

Recombinant polymerases for incorporation of protein shield nucleotide analogs

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing or nucleic acid amplification. Such properties include enhanced performance with large nucleotide analogs, increased stability, increased readlength, and improved detection of modified bases, and can also include resistance to photodamage, enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased accuracy, altered speed, increased cosolvent resistance, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

COOLED FAN FOR MICRO-CLIMATE CONTROL

A micro-climate control system includes a thermoelectric system integrated with a fan assembly. The thermoelectric system is operable to actively cool or heat air as the air passes through the fan assembly. The thermoelectric system includes a thermoelectric heat pump, a heat reject subsystem, and a heat accept subsystem. The fan assembly is operable to draw air from a space to be conditioned and output conditioned air passed through one of the heat reject subsystem and the heat accept subsystem to the space to be conditioned and output air passed through the other away from the space to be conditioned. In this way, the micro-climate control system may provide localized comfort, while allowing a larger climate control system to maintain a more efficient temperature set point. In this way, the overall energy consumption may be reduced while providing the same level of effective comfort. 1. A micro-climate control system , comprising:a fan assembly; and a thermoelectric heat pump;', 'a heat reject subsystem; and', 'a heat accept subsystem;, 'a thermoelectric system integrated with the fan assembly and being operable to actively cool or heat air as the air passes through the fan assembly, the thermoelectric system comprising draw air from a space to be conditioned;', 'output conditioned air passed through one of the heat reject subsystem and the heat accept subsystem to the space to be conditioned; and', 'output air passed through the other one of the heat reject subsystem and the heat accept subsystem away from the space to be conditioned., 'wherein the fan assembly is operable to2. The micro-climate control system of wherein:at least one of the heat reject subsystem and the heat accept subsystem comprises a radial heat exchanger.3. The micro-climate control system of wherein:the heat reject subsystem comprises the radial heat exchanger; andthe heat accept subsystem comprises the radial heat exchanger.4. The micro-climate control system of wherein:the thermoelectric ...

Context Aware Content Distribution

Described herein are systems and methods for context aware content distribution. Content such as e-books, presentations, audio, video, applications, and so forth may be distributed to user devices after establishing a trust relationship. An invitation to provide content may be sent to a user device based on the context of the user device, such as proximity to pre-determined devices, location, and so forth. Upon acceptance of the invitation to opt-in to receiving content, a trust relationship may be established which provides for the transfer of content to the user device. 1. (canceled)2. A method comprising:generating, by a server computer, an invitation message comprising a request to establish a first trust relationship between a user device and a first content server;sending the invitation message to the user device;receiving, by the server computer, an indication of acceptance from the user device;establishing the first trust relationship between the user device and the first content server;determining that the first content server comprises first content associated with a first geographic location;determining that a second content server comprises second content associated with the first geographic location;determining that a third content server comprises third content associated with the first geographic location;receiving, by the server computer, first device location information indicating a geographic location of the user device;determining that the geographic location is at or within a predetermined distance of the first geographic location;sending, based at least in part on the first trust relationship, a first approval message to the first content server, the first approval message causing the first content server to send the first content to the user device;determining that a trust relationship is not established between the user device and the second content server;preventing the second content server from sending the second content to the user device ...

RECOMBINANT POLYMERASES WITH INCREASED PHOTOTOLERANCE

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template. 1. A composition comprising a Φ29-type (phi29-type) recombinant DNA polymerase , which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:2 , and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position K128 , a K132Q substitution , a K132S substitution , an H146D substitution , a Q180F substitution , a G194D substitution , a G194E substitution , an I198E substitution , a K203E substitution , and an A434N substitution , wherein identification of positions is relative to SEQ ID NO:2 , and wherein said polymerase exhibits polymerase activity.2. The composition of claim 1 , wherein the recombinant polymerase comprises a K128E substitution or a K128Q substitution claim 1 , wherein identification of positions is relative to SEQ ID NO:2.3. The composition of claim 1 , wherein the recombinant polymerase comprises one or more mutation or combination of mutations selected from the group consisting of an amino acid substitution at position 250 claim 1 , an amino acid substitution at position 372 claim 1 , an amino acid substitution ...

SYSTEMS AND METHODS FOR HEIGHT CONTROL IN LASER METAL DEPOSITION

Disclosed is a welding system configured to perform additive manufacturing, particularly a welding system to achieve a stable a laser metal deposition with hot wire process by controlling the contact point between the welding wire and the workpiece. For example, the resistance of the stick-out wire is measured and the measured resistance is converted to a distance signal, which can then be used for comparison to a desired distance. The distance between the contact tip and the workpiece can then be adjusted based on the comparison. The present disclosure also relates to using a constant enthalpy system to determine and control the contact tip to workpiece distance. 1. An additive manufacturing system , comprising:an additive manufacturing tool configured to advance an electrode wire to a weld puddle on a surface of a workpiece;a heater configured to preheat the electrode wire at a location in an electrode wire feed path that is prior to the surface;a laser generator to provide a laser beam to create and heat the weld puddle to at least a melting temperature of the electrode wire; and monitor a current or a voltage associated with the electrode wire;', 'determine a resistance of the electrode wire based in part on the current or the voltage;', 'determine a distance between the tool and the surface based in part on the resistance; and', 'control a position of the tool to adjust the distance between the tool and the surface based on the resistance., 'a control circuitry configured to2. The system of claim 1 , the control circuitry further configured to:compare the distance against one or more threshold distances; andcontrol the position of the tool based the comparison.3. The system of claim 2 , wherein the one or more threshold distances are based on a predetermined range of angles measured between the electrode wire and the surface of the workpiece.4. The system of claim 3 , wherein the predetermined range of angles includes a 45-degree angle.5. The system of claim 3 ...

Laser intensity calibration

Systems, apparatuses, and methods are described for calibrating a laser power are described. A system may include a sample to be tested and a control sample that comprises a control analyte. A user may indicate a known concentration of the control analyte to the system (e.g., by entering a concentration value into a user interface or other process). The system may perform multiple runs at different laser powers and compare the measurements of each run against expected values for the control analyte at the known concentration. From that comparison, a calibrated laser power may be computed and that computed power level can be used by the system for the running of tests on an unknown sample.

Apparatus for neurosurgical stereotactic procedures

An apparatus and method are disclosed for displaying a path between a selected target and selected points on a patient's skull and for guiding surgical instruments along any selected path. The system is comprised of an image display system, an articulated arm and probe, and a stereotactic system. The sub-systems are coupled to one another so that the articulated probe may be used to select patient fiducial points that correspond to selected image fiducial points. Using these points, the image display system coregisters the external locations to the displayed images so that the probe condition may be displayed with the displayed images. The system further permits the identification of a selected target within a patient's brain and to project a path from the external position to the target prior to the performance of a craniotomy. After evaluation of the path, a surgeon may lock the stereotactic system in place to preserve a selected surgical path and to guide instruments along that path. A method of utilizing the system to perform such surgical procedures is also described.

Apparatus and method for surgical stereotactic procedures

An apparatus and method are disclosed for displaying a path between a selected target and selected points on a patient's skull and for guiding surgical instruments along any selected path. The system is comprised of an image display system, an articulated arm and probe, and a stereotactic system. The sub-systems are coupled to one another so that the articulated probe may be used to select patient fiducial points that correspond to selected image fiducial points. Using these points, the image display system coregisters the external locations to the displayed images so that the probe condition may be displayed with the displayed images. The system further permits the identification of a selected target within a patient's brain and to project a path from the external position to the target prior to the performance of a craniotomy. After evaluation of the path, a surgeon may lock the stereotactic system in place to preserve a selected surgical path and to guide instruments along that path. A method of utilizing the system to perform such surgical procedures is also described.

Metalworking system with force controlled wire feed start operation.

Un método para controlar un inicio de una operación de trabajo de metales. El método incluye detectar un contacto inicial entre un alambre que se alimenta desde un aparato de soldadura y una pieza de trabajo y, en respuesta a la detección, detener la alimentación del alambre desde el aparato de soldadura. El método incluye además activar una fuente de calor de alta energía configurada para calentar una punta del alambre y reanudar la alimentación del alambre desde el aparato de soldadura cuando la punta del alambre sea calentada por la fuente de calor de alta energía hasta un estado plástico. La alimentación del alambre se reanuda midiendo una retroalimentación de fuerza desde el alambre que entra en contacto con la pieza de trabajo. También se describe un aparato para implementar el método.

Wide path welding, cladding, additive manufacturing

A welding or cladding apparatus in which one or more energy beam emitters are used to generate a wide beam spot transverse to a welding or cladding path, and one or more wide feeders feed wire to the spot to create a wide welding or cladding puddle.

Acoustic perfusion devices

Acoustic perfusion devices for separating biological cells from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn.

Heat exchange fluid purification for dialysis system

Disclosed herein are small, lightweight, portable, systems that have the capability of reliably, reproducibly, highly efficiently and relatively inexpensively providing a source of purified water of sufficient volumes for home dialysis. In addition, the systems disclosed herein require much less purified water at any one time than the volumes typically needed for dialysis today, thereby further reducing the expense of running the system at home.

Laser welding, cladding, and/or additive manufacturing systems and methods of laser welding, cladding, and/or additive manufacturing

Systems and methods of a laser welding device are disclosed. The laser welding device includes a laser generator (102) configured to generate welding-type lasing power. A lens (104) focuses the welding-type lasing power at a focal point on a workpiece to generate a puddle during a welding-type operation. A wire feeder (123) is configured to feed wire to the puddle generated by the laser generator. A laser scanner (106) controls the lens to move the focal point of the welding-type lasing power in multiple dimensions over the workpiece during the welding-type operation. In some examples, the feed wire is used in an additive manufacturing process.

Acoustic perfusion devices

Methods are disclosed for separating beads and cells from a host fluid. The method includes flowing a mixture containing the host fluid, the beads, and the cells through an acoustophoretic device having an ultrasonic transducer including a piezoelectric material driven by a drive signal to create a multi-dimensional acoustic standing wave. A drive signal is sent to drive the at least one ultrasonic transducer to create the multi-dimensional acoustic standing wave. A recirculating fluid stream having a tangential flow path is located substantially tangential to the standing wave and separated therefrom by an interface region. A portion of the cells pass through the standing wave, and the beads are held back from the standing wave in the recirculating fluid stream at the interface region. Also disclosed is an acoustophoretic device having a coolant inlet adapted to permit the ingress of a cooling fluid into the device for cooling the transducer.

Wide path welding, cladding, additive manufacturing

A welding or cladding apparatus in which one or more energy beam emitters are used to generate a wide beam spot transverse to a welding or cladding path, and one or more wide feeders feed wire to the spot to create a wide welding or cladding puddle.

Methods and compositions for delivery of molecules and complexes to reaction sites

Hybrid additive manufacturing system using laser and arc welding

Detection of unauthorized information exchange between users

Detection and prevention of unauthorized exchange of information between users of user devices is disclosed. Monitoring data associated with respective information received by the user devices from respective users may be received. Position information indicating proximity or relative positions of the devices may also be received. The monitoring data and/or the position information may be analyzed to determining whether unauthorized information exchange between users may be occurring. One or more metrics indicating a likelihood that unauthorized information exchange is occurring may be generated based on the analysis of the monitoring data and/or the position information. A generated metric may be compared to an associated threshold and preventive measures for deterring unauthorized information exchange may be identified based on a result of the comparison.

Systems and methods for adaptive control of wire preheating

A welding-type system includes a power supply configured to control preheating of an electrode wire. A controller is configured to receive a plurality of power values corresponding to a power output of the power supply and calculate an arc power value corresponding to an arc condition at the preheated electrode wire based on a rate of change of the plurality of power values. A target power output value is determined based on the calculated arc power value, and the power output is adjusted based on the determined target power value.

Laser welding, cladding, and/or additive manufacturing systems and methods of laser welding, cladding, and/or additive manufacturing.

Se divulgan sistemas y métodos de un dispositivo de soldadura láser. El dispositivo de soldadura láser incluye un generador láser configurado para generar energía láser de tipo soldadura. Una lente concentra la energía láser de tipo soldadura en un punto focal sobre una pieza de trabajo para generar un baño de fusión durante una operación de tipo soldadura. Un alimentador de alambre se configura para suministrar alambre al baño de fusión que se genera mediante el generador láser. Un escáner láser controla la lente para mover el punto focal de la energía láser de tipo soldadura en múltiples dimensiones sobre la pieza de trabajo durante la operación de tipo soldadura. En algunos ejemplos, el alambre suministrado se utiliza en un proceso de fabricación aditiva.

Dispensing control systems and methods

Sistema de dispensación (100) para entregar un material a un componente de recepción situado corriente abajo delsistema de dispensación, comprendiendo el sistema de dispensación :un receptáculo (110) configurado para mezclar el material y agua;una válvula (145);un dispositivo de medición de material (515) configurado para dispensar el material en el receptáculo;un sensor (525) situado próximo al receptáculo y configurado para generar una primera señal indicativa deconductividad de solución de agua y/o material; yun controlador (505),caracterizado por el hecho de que la válvula se configura para controlar un suministro de agua al receptáculo,teniendo la válvula una posición de cerrado que impide que entre agua en el receptáculo y una posición de abierto quepermite que entre agua en el receptáculo y el controlador es configurado para recibir la primera señal del sensor y paragenerar una señal de control de válvula y una señal de control del dispositivo de medición del material, siendo la señalde control de válvula operable para alternar la válvula entre la posición de abierto y la posición de cerrado, siendo laseñal de control del dispositivo de medición de material operable para iniciar una dispensación del material, siendogeneradas la señal de control de válvula y la señal de dispositivo de medición del material al menos parcialmente enrespuesta a una comparación por el controlador de la primera señal con uno o más valores umbral de conductividadalmacenados. Dispensing system (100) for delivering a material to a receiving component located downstream of the dispensing system, the dispensing system comprising: a receptacle (110) configured to mix the material and water; a valve (145); a device for measurement of material (515) configured to dispense the material in the receptacle; a sensor (525) located next to the receptacle and configured to generate a first indicative signal of water and / or material solution conductivity; and a controller (505), characterized in ...

Delaying real-time sequencing

Methods, compositions, and systems are provided that allow for reliable sequencing of the initial sequence region of a sequence of interest. The methods of the invention allow for more reliable barcoding of subpopulations of nucleic acids to be sequenced.

Coaxial laser hotwire head

A laser processing head includes first (116) and second (118) contact points connected to a power source. The power source generates a current to flow through an electrode wire (112) between the first and second contact points to heat the electrode wire. A laser source generates one or more laser beams (108A, 108B) having lasing power sufficient to at least partially melt the electrode wire. A coaxial laser head focuses the one or more laser beams at one or more focal points (110) on a workpiece (114) to at least partially melt the electrode wire.