DROPLET OPERATIONS DEVICE

The invention provides droplet actuators with droplet operations surfaces for manipulating droplets, e.g., by conducting droplet operations. The droplet operations surfaces are typically exposed to a droplet operations gap. One or more regions of a droplet operation surface may include patterned topographic features. The invention also provides a droplet actuator in which one or both gap-facing droplet operations surfaces is formed using a removable film. The removable film may, in various embodiments, also include other components ordinarily associated with the droplet actuator substrate, such as the dielectric layer and the electrodes. Further, the invention provides droplet actuator devices and methods for coupling and/or sealing substrates of a droplet actuator, such as techniques for self-aligning assembly of droplet actuator substrates. The invention provides droplet actuators and methods of disassembling the droplet actuator in order to provide access for cleaning and/or recycling of droplet actuator surfaces. 1242-. (canceled)243. A droplet actuator comprising:(a) a substantially disk-shaped droplet operations substrate;(b) droplet operations electrodes arranged in radially oriented paths on the substrate to provide wedge-shaped unit cells, wherein the droplet operations electrodes are configured for conducting droplet operations on a surface of the substrate;(c) one or more reservoirs associated with the wedge-shaped unit cells; and(d) contact pads associated with the wedge-shaped unit cells and configured for providing electrical connections to the droplet operations electrodes arranged thereon.244. The droplet actuator of claim 243 , wherein the wedge-shaped unit cells are configured to operate independently of one another.245. The droplet actuator of claim 243 , wherein the one or more reservoirs comprise one or more sample reservoirs claim 243 , reagent reservoirs claim 243 , wash reservoirs claim 243 , and/or waste reservoirs.246. The droplet actuator ...

Bead Incubation and Washing on a Droplet Actuator

The invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay.

Bead Manipulation Techniques

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads. 146-. (canceled)47. An apparatus comprising: an electrowetting (EW) configuration comprising a droplet operations surface; and', 'a dielectrophoresis (DEP) configuration comprising a surface,', 'wherein the DEP configuration is disposed adjacent to the EW configuration such that the surface of the DEP configuration is adjacent to the droplet operations surface., 'a first substrate comprising48. The apparatus of wherein the droplet operations surface comprises one or more electrodes arranged for conducting one or more droplet operations on the surface.49. The apparatus of wherein the one or more electrodes comprise electrowetting electrodes.50. The apparatus of wherein the one or more electrodes comprise a path of electrowetting electrodes.51. The apparatus of further comprising a dielectric layer atop the one or more electrodes.52. The apparatus of wherein the DEP configuration comprises a DEP electrode configuration comprising one or more electrodes.53. The apparatus of wherein the DEP electrode configuration is arranged for attracting and/ ...

Droplet actuator with improved top substrate

The invention provides a droplet actuator. The droplet actuator may include a base substrate and a top substrate separated to form a gap. The base substrate may include electrodes configured for conducting droplet operations in the gap; and the top substrate may include a glass substrate portion coupled to a non-glass portion, where the non-glass portion may include one or more openings establishing a fluid path extending from an exterior of the droplet actuator and into the gap. The invention also provides related methods of manufacturing the droplet actuator, methods of using the droplet actuator, and methods of loading the droplet actuator.

Microfluidic feedback using impedance detection

Methods comprising measuring the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode are disclosed. Computer readable mediums storing processor executable instructions for performing the method, and systems are also disclosed. The systems comprise a processor, memory and code stored in the memory that when executed cause the processor at least to: receive an output voltage signal, superimpose an excitation signal onto the output voltage signal to produce a superimposed signal, connect the superimposed signal to an electrode in a droplet actuator, suppress the output voltage signal, when detecting an impedance of the electrode, and measure the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode.

Droplet actuator configurations and methods of conducting droplet operations

A droplet actuator with a droplet formation electrode configuration associated with a droplet operations surface, wherein the electrode configuration may include one or more electrodes configured to control volume of a droplet during formation of a sub-droplet on the droplet operations surface. Methods of making and using the droplet actuator are also provided.

Method of concentrating beads in a droplet

Methods of concentrating beads in a droplet and/or loading beads on a fluidic device are provided, including among other things, a method of concentrating beads in a droplet, the method comprising: (a) providing a droplet actuator comprising: (i) an interior droplet operations volume; and (ii) a reservoir exterior to the interior volume; (iii) a droplet established in a liquid path extending from the reservoir into the interior volume; (b) providing magnetically responsive beads in the portion of the droplet which is in the reservoir; (c) magnetically attracting the magnetically responsive beads through the liquid path into the portion of the droplet which is in the interior volume; and (d) forming a droplet comprising one or more of the magnetically responsive beads in the interior volume.

Droplet Operations Device

The invention provides droplet actuators with droplet operations surfaces for manipulating droplets, e.g., by conducting droplet operations. The droplet operations surfaces are typically exposed to a droplet operations gap. One or more regions of a droplet operation surface may include patterned topographic features. The invention also provides a droplet actuator in which one or both gap-facing droplet operations surfaces is formed using a removable film. The removable film may, in various embodiments, also include other components ordinarily associated with the droplet actuator substrate, such as the dielectric layer and the electrodes. Further, the invention provides droplet actuator devices and methods for coupling and/or sealing substrates of a droplet actuator, such as techniques for self-aligning assembly of droplet actuator substrates. The invention provides droplet actuators and methods of disassembling the droplet actuator in order to provide access for cleaning and/or recycling of droplet actuator surfaces. 1. A droplet actuator substrate comprising:(a) a base substrate comprising electrodes;(b) an adhesive layer atop the base substrate;(c) a dielectric layer atop the adhesive layer and bound to the base substrate by the adhesive layer; and(d) a droplet operations surface atop the dielectric layer.2. The droplet actuator substrate of wherein the droplet operations surface has a three dimensional topology which differs from a three dimensional topology of the base substrate comprising electrodes.3. The droplet actuator substrate of wherein the adhesive layer and dielectric layer establish a droplet operations topology selected to enhance one or more droplet operations.4. The droplet actuator substrate of further comprising a hydrophobic layer atop the dielectric layer.5. The droplet actuator substrate of wherein the adhesive layer is selected to flow under heat and pressure claim 1 , where the heat and pressure required to cause such flow are sufficiently ...

Droplet Actuator and Droplet-Based Techniques

The invention is directed to certain droplet actuated molecular techniques. In one embodiment, the invention provides droplet actuator methods for detection of single nucleotide polymorphisms (SNPs) in a DNA sequence using digital microfluidics, including droplet actuator-based sample preparation and SNP analysis. In another embodiment, the invention provides droplet actuator devices and methods for providing integrated sample preparation and multiplexed detection of an infectious agent, such as HIV. In yet another embodiment, the invention provides droplet actuator devices and techniques for PCR amplification and detection of specific nucleic acid sequences using digital microfluidics, including droplet actuator-based sample preparation and target nucleic acid analysis. In yet another embodiment the invention provides methods for performing hot-start PCR on a droplet actuator. In yet another embodiment, the method of the invention combines PCR amplification with pyrosequencing to investigate specific sequences. 1137-. (canceled)138. A method of optimizing real-time PCR , the method comprising:(a) providing a digital microfluidics system comprising at least one signal monitor;(b) providing a sample droplet comprising a detectable signal component;(c) conducting a PCR reaction;(d) monitoring intensity of the detectable signal component during thermal cycles; and(e) initiating a next thermal cycle when the signaling component of a sample reaches a plateau.139. The method of claim 138 , wherein initiating a next thermal cycle is automated such that when the signaling component of a sample reaches a plateau the next thermal cycle is automatically initiated.140. The method of claim 138 , wherein the thermal cycle comprises annealing.141. The method of claim 138 , wherein the thermal cycle comprises extension.142. The method of claim 138 , wherein the detectable signal comprises fluorescence.143. The method of claim 142 , wherein the fluorescence comprises EVA GREEN dye. ...

Method of Loading a Droplet Actuator

A method of loading a droplet actuator is provided. In one embodiment, the method may include, providing: a droplet actuator loaded with a filler fluid; a reservoir comprising a droplet fluid; and a fluid path extending from the reservoir into the droplet actuator; and forcing filler fluid from one locus in the droplet actuator to another locus in the droplet actuator; or out of the droplet actuator; thereby causing droplet fluid to flow through the fluid path and into the droplet actuator. 1. A method of loading a droplet actuator , the method comprising: (i) a droplet actuator loaded with a filler fluid;', '(ii) a reservoir comprising a droplet fluid; and', '(iii) a fluid path extending from the reservoir into the droplet actuator; and, '(a) providing (i) from one locus in the droplet actuator to another locus in the droplet actuator; or', '(ii) out of the droplet actuator;, '(b) forcing filler fluidthereby causing droplet fluid to flow through the fluid path and into the droplet actuator.2. The method of wherein step (b) comprises forcing droplet fluid into sufficient proximity with one or more electrodes to enable one or more droplet operations to be mediated in the droplet actuator by the one or more electrodes.3. The method of wherein step (b) comprises forcing the filler fluid using a negative pressure source.4. The method of wherein step (b) comprises forcing the filler fluid using a positive pressure source.5. The method of wherein multiple droplet fluids are loaded from multiple reservoirs.6. The method of wherein the droplet operation comprises a droplet dispensing operation in which a droplet is dispensed from the droplet fluid. In addition to the patent applications cited herein, each of which is incorporated herein by reference, this application is a divisional of U.S. patent application Ser. No. 12/747,231, filed Aug. 25, 2010, entitled “Droplet Actuator Configurations and Methods”, the application of which is a national phase application of PCT/ ...

Droplet Actuator

Droplet actuator for conducting droplet operations, such as droplet transport and droplet dispensing, is provided. In one embodiment, the droplet actuator may include an electrode that is rotationally but not reflectively symmetrical. 1. A droplet actuator comprising an electrode that is rotationally but not reflectively symmetrical.2. The droplet actuator of comprising a path and/or array of the electrodes.3. The droplet actuator of wherein the electrodes are interdigitated.4. The droplet actuator of wherein the electrodes are not interdigitated.5. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is 3.6. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is 4.7. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is 5.8. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is 6.9. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is 7.10. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is 8.11. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is 9.12. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is 10.13. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and X is greater than 10.14. The droplet actuator of wherein the rotational symmetry is X-fold claim 1 , and for each electrode X is 3 claim 1 , 4 claim 1 , 5 claim 1 , 6 claim 1 , 7 claim 1 , 8 claim 1 , 9 claim 1 , and/or 10.15. The droplet actuator of wherein the droplet actuator comprises a path and/or array of the electrodes claim 1 , wherein adjacent electrodes are arranged such that no line can be drawn between two adjacent electrodes without overlapping one or both of the two adjacent electrodes.16. The droplet actuator of wherein the electrodes are not interdigitated.17. The droplet actuator of wherein the electrodes are ...

Method of Reducing Liquid Volume Surrounding Beads

The invention provides droplet actuators and droplet actuator techniques. Among other things, the droplet actuators and methods are useful for manipulating beads on a droplet actuator, such as conducting droplet operations using bead-containing droplets on a droplet actuator. For example, beads may be manipulated on a droplet actuator in the context of executing a sample preparation protocol and/or an assay protocol. An output of the methods of the invention may be beads prepared for execution of an assay protocol. Another output of the methods of the invention may be results of an assay protocol executed using beads. Among the methods described herein are methods of concentrating beads in droplets, methods of washing beads, methods of suspending beads, methods of separating beads, methods of localizing beads within a droplet, methods of forming emulsions in which droplets include beads, methods of loading beads into a droplet operations gap of a droplet actuator, methods of organizing beads in a monolayer, and methods of capturing, trapping or restraining beads. 1. A method of reducing a liquid volume surrounding one or more magnetically responsive beads , the method comprising:(a) providing a droplet comprising the one or more magnetically responsive beads;(b) exposing the one or more magnetically responsive beads in the droplet to a first region of a magnetic field; and(c) separating the droplet from the first region of the magnetic field, the one or more magnetically responsive beads remaining in the magnetic field.2. The method of further comprising heating the droplet to a temperature which permits the one or more magnetically responsive beads to break away from the droplet.3. The method of further comprising adjusting the droplet to a shape which permits the one or more magnetically responsive beads to break away from the droplet.4. The method of further comprising adjusting the droplet to an interfacial tension which permits the one or more magnetically ...

BEAD INCUBATION AND WASHING ON A DROPLET ACTUATOR

The present invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay. 1. A method of manipulating a droplet comprising magnetically responsive beads therein , the method comprising: (i) droplet operations electrodes arranged for conducting droplet operations on a droplet operations surface;', '(ii) the droplet, arranged on the droplet operations surface and subject to droplet operations mediated by the droplet operations electrodes; and', '(ii) a magnet having a magnetic field and positioned relative to the droplet operations surface such that the droplet may be transported into the magnetic field of the magnet and transported out of the magnetic field of the magnet;, '(a) providing a droplet actuator comprising(b) transporting the droplet into the magnetic field of the magnet by executing droplet operations mediated by the droplet operations electrodes;(c) transporting the droplet out of the magnetic field of the magnet executing droplet operations mediated by the droplet operations electrodes without removing the magnet.2. The method of claim 1 , wherein the droplet transported has a footprint of from about one to about three times the area of a single droplet operations electrode.3. The method of claim 1 , wherein the droplet transported has a footprint of from about two to about three times the area of a single droplet operations ...

BEAD MANIPULATION TECHNIQUES

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads. 121-. (canceled)22. A method of washing beads , the method comprising:(a) providing the beads in a sample droplet comprising a target substance on a droplet operations substrate within a magnetic field, the droplet operations substrate comprising a path of electrowetting electrodes;(b) transporting the sample droplet away from the beads by sequentially activating the electrowetting electrodes, causing the beads to be snapped off as the droplet is transported away, yielding a supernatant droplet and leaving behind a daughter droplet comprising substantially all of the magnetically responsive beads; and(c) subjecting the daughter droplet to a merge-and-split bead washing protocol.23. The method of wherein the supernatant droplet includes more than 50% of the unbound substances being removed.24. The method of wherein the supernatant droplet includes more than 75% of the unbound substances being removed.25. The method of further comprising:(a) merging the daughter droplet comprising substantially all of the magnetically responsive beads with a ...

Bead incubation and washing on a droplet actuator

Methods are provided for separating magnetically responsive beads from a droplet in a droplet actuator. Droplet operations electrodes and a magnet are arranged in a droplet actuator to manipulate a bead-containing droplet and position it relative to a magnetic field region that attracts the magnetically responsive beads. The droplet operations electrodes are operated to control the droplet shape and transport it away from the magnetic field region to form a concentration of beads in the droplet. The continued transport of the droplet away from the magnetic field causes the concentration of beads to break away from the droplet to yield a small, concentrated bead-containing droplet immobilized by the magnet.

BEAD INCUBATION AND WASHING ON A DROPLET ACTUATOR

The present invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay. 19-. (canceled)10. A method of resuspending magnetically responsive beads between wash cycles , the method comprising: (i) droplet operations electrodes arranged for conducting droplet operations on a droplet operations surface; and', "(ii) a magnet positioned relative to the droplet operations surface such that a droplet controlled by one or more of the droplet operations electrodes may be positioned within or away from a first region of the magnet's magnetic field capable of substantially attracting magnetically responsive beads in the droplet;"], '(a) providing a droplet actuator comprising(b) positioning a droplet having magnetically responsive beads therein at a location partially overlapping the first region of the magnetic field;(c) transporting the droplet through activation of selected droplet operations electrodes away from the first region of the magnetic field;(d) operating the droplet operations electrodes to cause the droplet to move towards the first region of the magnetic field; and(e) repeating steps (c) and (d) to cause sufficient resuspension of beads such that unbound material may be effectively removed in subsequent wash cycles.11. The method of claim 10 , wherein step (b) is conducted by positioning the droplet at a location partially overlapping a ...

Bead Manipulation Techniques

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads.

Droplet operations device

The invention provides droplet actuators with droplet operations surfaces for manipulating droplets, e.g., by conducting droplet operations. The droplet operations surfaces are typically exposed to a droplet operations gap. One or more regions of a droplet operation surface may include patterned topographic features. The invention also provides a droplet actuator in which one or both gap-facing droplet operations surfaces is formed using a removable film. The removable film may, in various embodiments, also include other components ordinarily associated with the droplet actuator substrate, such as the dielectric layer and the electrodes. Further, the invention provides droplet actuator devices and methods for coupling and/or sealing substrates of a droplet actuator, such as techniques for self-aligning assembly of droplet actuator substrates. The invention provides droplet actuators and methods of disassembling the droplet actuator in order to provide access for cleaning and/or recycling of droplet actuator surfaces.

Bead Manipulations on a Droplet Actuator

A droplet actuator comprising: (a) a base substrate comprising electrodes configured for conducting droplet operations on a droplet operations surface thereof; (b) a droplet comprising one or more beads situated on the droplet operations surface; (c) a barrier arranged in relation to the droplet and the electrodes such that a droplet may be transported away from the beads using one or more droplet operations mediated by one or more of the electrodes while transport of the beads is restrained by a barrier. Related methods and kits are also provided 1. A droplet actuator comprising:(a) a base substrate comprising electrodes configured for conducting droplet operations on a droplet operations surface thereof;(b) a droplet comprising one or more beads situated on the droplet operations surface;(c) a barrier arranged in relation to the droplet and the electrodes such that a droplet may be transported away from the beads using one or more droplet operations mediated by one or more of the electrodes while transport of the beads is restrained by a barrier.2. The droplet actuator of further comprising a top substrate separated from the droplet operations surface to form a gap for conducting droplet operations.3. The droplet actuator of wherein the barrier is coupled to and extends downward from the top substrate.4. The droplet actuator of wherein the barrier is configured to leave a gap between a bottom edge of the barrier and the droplet operations surface.5. The droplet actuator of wherein the barrier comprises a vertical gap through which fluid may pass during a droplet operation mediated by one or more of the electrodes.6. The droplet actuator of wherein the vertical gap is situated over an electrode.7. The droplet actuator of wherein the vertical gap extends substantially from a surface of the top substrate facing the gap and the droplet operations surface.8. The droplet actuator of wherein the one or more beads are completely surrounded by the barrier.9. The droplet ...

Droplet Actuator Devices and Methods for Manipulating Beads

The invention provides droplet actuators and droplet actuator techniques. Among other things, the droplet actuators and methods are useful for manipulating beads on a droplet actuator, such as conducting droplet operations using bead-containing droplets on a droplet actuator. For example, beads may be manipulated on a droplet actuator in the context of executing a sample preparation protocol and/or an assay protocol. An output of the methods of the invention may be beads prepared for execution of an assay protocol. Another output of the methods of the invention may be results of an assay protocol executed using beads. Among the methods described herein are methods of concentrating beads in droplets, methods of washing beads, methods of suspending beads, methods of separating beads, methods of localizing beads within a droplet, methods of forming emulsions in which droplets include beads, methods of loading beads into a droplet operations gap of a droplet actuator, methods of organizing beads in a monolayer, and methods of capturing, trapping or restraining beads. 1232-. (canceled)233. A method of positioning a droplet comprising magnetically responsive beads atop an electrode in a droplet operations gap of a droplet actuator , the method comprising using a magnetic field to attract the droplet comprising the beads to a position atop an electrode , wherein the droplet has a droplet footprint and the electrode has an electrode footprint , and wherein the droplet footprint area is substantially smaller than the electrode footprint area.234. The method of wherein the droplet footprint area is less than about 75% of the electrode footprint area.235. The method of wherein the droplet footprint area is less than about 50% of the electrode footprint area.236. The method of wherein the droplet footprint area is less than about 25% of the electrode footprint area.237. The method of wherein the droplet is substantially surrounded in the droplet operations gap by a liquid ...

MICROFLUIDIC FEEDBACK USING IMPEDANCE DETECTION

Methods comprising measuring the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode are disclosed. Computer readable mediums storing processor executable instructions for performing the method, and systems are also disclosed. The systems comprise a processor, memory and code stored in the memory that when executed cause the processor at least to: receive an output voltage signal, superimpose an excitation signal onto the output voltage signal to produce a superimposed signal, connect the superimposed signal to an electrode in a droplet actuator, suppress the output voltage signal, when detecting an impedance of the electrode, and measure the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode. 1. A method , comprising:(a) receiving an output voltage signal produced by a power supply;(b) superimposing an excitation signal onto the output voltage signal to produce a superimposed signal;(c) connecting the superimposed signal to an electrode in a droplet actuator;(d) suppressing the output voltage signal when detecting an impedance of the electrode; and(e) measuring the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode.2. The method of claim 1 , wherein superimposing the excitation signal comprises adding the excitation signal to the output voltage signal.3. The method of claim 1 , wherein suppressing the output voltage signal comprises stopping the output voltage signal.4. The method of claim 1 , wherein suppressing the output voltage signal comprises disabling a switching action of the power supply.5. The method of claim 1 , wherein suppressing the output voltage signal comprises disabling the power supply.6. The method of claim 1 , further comprising receiving a suppression signal to suppress the output voltage signal from the power ...

BEAD MANIPULATION TECHNIQUES

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include providing a droplet including magnetically responsive beads. The droplet may be provided within a region of a magnetic field having sufficient strength to attract the magnetically responsive beads to an edge of the droplet or towards an edge of the droplet, or otherwise regionalize or aggregate beads within the droplet. The method may also include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads. 1. A method of redistributing magnetically responsive beads in a droplet , the method comprising:(a) providing a droplet comprising magnetically responsive beads within a region of a magnetic field having sufficient strength to attract the magnetically responsive beads to an edge of the droplet; and(b) using electrodes to conduct on a droplet operations surface a droplet operation using the droplet without removing the magnetically responsive beads from the region of the magnetic field, thereby redistributing the magnetically responsive beads within the droplet.2. The method of wherein the droplet comprises a sample droplet comprising a target analyte.3. The method of wherein the redistributing of ...

Droplet Actuator with Improved Top Substrate

The invention provides a droplet actuator. The droplet actuator may include a base substrate and a top substrate separated to form a gap. The base substrate may include electrodes configured for conducting droplet operations in the gap; and the top substrate may include a glass substrate portion coupled to a non-glass portion, where the non-glass portion may include one or more openings establishing a fluid path extending from an exterior of the droplet actuator and into the gap. The invention also provides related methods of manufacturing the droplet actuator, methods of using the droplet actuator, and methods of loading the droplet actuator. 1. A droplet actuator comprising a base substrate and a top substrate separated to form a gap , wherein:(a) the base substrate comprises electrodes configured for conducting droplet operations in the gap; and(b) the top substrate comprises a glass substrate portion coupled to a non-glass portion, where the non-glass portion comprises one or more openings establishing a fluid path extending from an exterior of the droplet actuator and into the gap.2. The droplet actuator of wherein the non-glass portion comprises a plastic or resin portion.3. The droplet actuator of wherein the non-glass portion comprises a portion into which the glass substrate portion is inserted.4. The droplet actuator of wherein the fluid path is arranged to flow fluid into an actual or virtual reservoir associated with one or more reservoir electrodes associated with the base substrate.5. The droplet actuator of wherein the fluid path is arranged to flow fluid into proximity with one or more of the electrodes.6. The droplet actuator of wherein the glass substrate portion does not include openings therein.7. The droplet actuator of wherein:(a) the non-glass portion overlaps the glass substrate portion; and(b) an aperture is provided in the non-glass portion for providing a sensing path from the gap, through the glass substrate portion, through the aperture ...

Droplet Actuator Configurations and Methods of Conducting Droplet Operations

A droplet actuator with a droplet formation electrode configuration associated with a droplet operations surface, wherein the electrode configuration comprises one or more electrodes configured to control volume of a droplet during formation of a sub-droplet on the droplet operations surface. Methods of making and using the droplet actuator are also provided.

BEAD INCUBATION AND WASHING ON A DROPLET ACTUATOR

Methods are provided for separating magnetically responsive beads from a droplet in a droplet actuator. Droplet operations electrodes and a magnet are arranged in a droplet actuator to manipulate a bead-containing droplet and position it relative to a magnetic field region that attracts the magnetically responsive beads. The droplet operations electrodes are operated to control the droplet shape and transport it away from the magnetic field region to form a concentration of beads in the droplet. The continued transport of the droplet away from the magnetic field causes the concentration of beads to break away from the droplet to yield a small, concentrated bead-containing droplet immobilized by the magnet. 1. A droplet actuator comprising:droplet operations electrodes to conduct droplet operations on a droplet operations surface; and(ii) a magnet positioned relative to the droplet operations surface such that a droplet controlled by one or more of the droplet operations electrodes may be positioned within or away from a first region of the magnet's magnetic field to attract magnetically responsive beads in the droplet;wherein the magnet is positioned such that three or more droplet operations electrodes are within the magnetic field of the magnet.2. The method of claim 1 , wherein the magnet is an electromagnet.3. The method of claim 1 , wherein the magnetically responsive beads are coated with a primary antibody with an affinity for a specific target antigen.4. A method comprising:positioning a droplet having magnetically response beads located therein away from a magnet of a droplet actuator, where the droplet actuator comprises droplet operations electrodes to conduct droplet operations on a droplet operations surface, and where the droplet actuator further comprises the magnet, where the magnet is positioned relative to the droplet operations surface such that a droplet controlled by one or more of the droplet operations electrodes may be positioned within or ...

BEAD MANIPULATION TECHNIQUES

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads. 121-. (canceled)22. A droplet actuator comprising: (i) a droplet operations surface; and', '(ii) electrodes arranged for conducting one or more electrowetting-mediated droplet operations on the surface; and, '(a) a first substrate comprising(b) one or more dielectrophoresis electrode configurations arranged for attracting and/or trapping one or more particles in a droplet situated on the droplet operations surface.23. The droplet actuator of wherein the droplet actuator comprises a second substrate separated from the droplet operations surface to form a droplet operations gap.24. The droplet actuator of wherein the one or more dielectrophoresis electrode configurations comprise at least one dielectrophoresis electrode configuration mounted on the second substrate.25. The droplet actuator of wherein the dielectrophoresis electrode configurations comprise at least one quadripole electrode configuration.26. The droplet actuator of wherein quadripole electrode configuration comprises four opposing triangular electrodes arranged to form a ...

DROPLET ACTUATOR AND DROPLET-BASED TECHNIQUES

The invention is directed to certain droplet actuated molecular techniques. In one embodiment, the invention provides droplet actuator methods for detection of single nucleotide polymorphisms (SNPs) in a DNA sequence using digital microfluidics, including droplet actuator-based sample preparation and SNP analysis. In another embodiment, the invention provides droplet actuator devices and methods for providing integrated sample preparation and multiplexed detection of an infectious agent, such as HIV. In yet another embodiment, the invention provides droplet actuator devices and techniques for PCR amplification and detection of specific nucleic acid sequences using digital microfluidics, including droplet actuator-based sample preparation and target nucleic acid analysis. In yet another embodiment the invention provides methods for performing hot-start PCR on a droplet actuator. In yet another embodiment, the method of the invention combines PCR amplification with pyrosequencing to investigate specific sequences. 1165-. (canceled)166. A method of detecting a signal in a droplet , comprising:(a) providing a droplet comprising a reagent producing a detectable signal;(b) aggregating the reagent in a region of the droplet; and(c) sensing to detect the detectable signal in the region of aggregation and/or in a region of the droplet in which the reagent is not aggregated.167. The method of wherein the region is at an edge of the droplet.168. The method of wherein the region is away from an edge of the droplet.169. The method of wherein the reagent is coupled directly or indirectly to beads.170. The method of wherein the beads are magnetically responsive beads and the aggregation is effected by a magnetic field.171. The method of claim 170 , wherein the region of aggregation is at an edge of the droplet.172. The method of claim 170 , wherein the magnetic field pulls a finger of droplet fluid from the droplet without splitting the droplet and the region of aggregation is at ...

Bead Incubation and Washing on a Droplet Actuator

The invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay. 19-. (canceled)10. A method of separating beads from a droplet onto a magnet , the method comprising: (i) droplet operations electrodes arranged for conducting droplet operations on a droplet operations surface; and', "(ii) a magnet positioned relative to the droplet operations surface such that a droplet controlled by one or more of the droplet operations electrodes may be positioned within or away from a first region of the magnet's magnetic field capable of substantially attracting magnetically responsive beads in the droplet;"], '(a) providing a droplet actuator comprising(b) positioning a droplet having magnetically responsive beads therein within the first region of the magnetic field of the magnet to cause the beads to be attracted to the magnet, and activating the droplet operations surface in a manner to cause the droplet to take an elongate shape;(c) operating the droplet operations surface to activate one electrode at a time, to cause the droplet to move away from the magnet, and thereby cause the geometry of the droplet to be distorted; and(d) continuing to operate the droplet operations surface to transport the droplet further away from the magnet and inactivating an electrode intermediate to the droplet to cause the droplet to split into a supernatant droplet and a ...

PLASMON RESONANCE (PR) SYSTEM AND INSTRUMENT, DIGITAL MICROFLUIDIC (DMF) CARTRIDGE, AND METHODS OF USING LOCALIZED SURFACE PLASMON RESONANCE (LSPR) FOR ANALYSIS OF ANALYTES

A plasmon resonance (PR) system and instrument, digital microfluidic (DMF) cartridge, and methods of using localized surface plasmon resonance (LSPR) and droplet operations for analysis of analytes is disclosed. For example, a PR system is provided that may include a DMF cartridge that may support both fixed LSPR sensing capability and in-solution LSPR sensing capability for analysis of analytes. The DMF cartridge may include an electrode arrangement for performing droplet operations, wherein the droplet operations can be used for performing fixed LSPR sensing operations and in-solution LSPR sensing operations. Further, methods of using droplet operations in the DMF cartridge to perform fixed LSPR sensing operations and/or in-solution LSPR sensing operations are provided. 1. A cartridge for use with an instrument to perform measurement of a fluid , comprising:a digital microfluidics (DMF) portion comprising a plurality of droplet actuators operative to perform droplet operations on a fluid droplet in the DMF portion; anda reaction portion comprising sensor media that is disposed in relation to the plurality of droplet actuators, wherein the plurality of droplet actuators are operative to induce movement of the fluid droplet relative to the sensor media while in contact with the sensor media.2. The cartridge of claim 1 , wherein the plurality of droplet actuators comprise a plurality of reaction electrodes.3. The cartridge of claim 2 , wherein the plurality of reaction electrodes perform droplet operations by electrowetting.4. The cartridge of claim 3 , wherein the sensor media comprises surface plasmon resonance (SPR) sensor media.5. The cartridge of claim 4 , wherein the SPR sensor media is functionalized with a capture molecule to which a target molecule of an analyte fluid binds to change an optical signal of the SPR sensor media.6. The cartridge of claim 5 , wherein the capture molecule comprises a ligand immobilized on the SPR sensor media that is sensitive to ...

METHOD OF CONCENTRATING BEADS IN A DROPLET

Methods of concentrating beads in a droplet and/or loading beads on a fluidic device are provided, including among other things, a method of concentrating beads in a droplet, the method comprising: (a) providing a droplet actuator comprising: (i) an interior droplet operations volume; and (ii) a reservoir exterior to the interior volume; (iii) a droplet established in a liquid path extending from the reservoir into the interior volume; (b) providing magnetically responsive beads in the portion of the droplet which is in the reservoir; (c) magnetically attracting the magnetically responsive beads through the liquid path into the portion of the droplet which is in the interior volume; and (d) forming a droplet comprising one or more of the magnetically responsive beads in the interior volume.

Plasmon resonance (PR) system and instrument, digital microfluidic (DMF) cartridge, and methods of using localized surface plasmon resonance (LSPR) for analysis of analytes

A plasmon resonance (PR) system and instrument, digital microfluidic (DMF) cartridge, and methods of using localized surface plasmon resonance (LSPR) and droplet operations for analysis of analytes is disclosed. For example, a PR system is provided that may include a DMF cartridge that may support both fixed LSPR sensing capability and in-solution LSPR sensing capability for analysis of analytes. The DMF cartridge may include an electrode arrangement for performing droplet operations, wherein the droplet operations can be used for performing fixed LSPR sensing operations and in-solution LSPR sensing operations. Further, methods of using droplet operations in the DMF cartridge to perform fixed LSPR sensing operations and/or in-solution LSPR sensing operations are provided.

Bead manipulation techniques

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads.

Bead incubation and washing on a droplet actuator

The invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay.

Reagent and sample preparation and loading on a fluidic device

Methods of concentrating beads in a droplet and/or loading beads on a fluidic device are provided, including among other things, a method of concentrating beads in a droplet, the method comprising: (a) providing a droplet actuator comprising: (i) an interior droplet operations volume; and (ii) a reservoir exterior to the interior volume; (iii) a droplet established in a liquid path extending from the reservoir into the interior volume; (b) providing magnetically responsive beads in the portion of the droplet which is in the reservoir; (c) magnetically attracting the magnetically responsive beads through the liquid path into the portion of the droplet which is in the interior volume; and (d) forming a droplet comprising one or more of the magnetically responsive beads in the interior volume.

Bead incubation and washing on a droplet actuator

Methods are provided for separating magnetically responsive beads from a droplet in a droplet actuator. Droplet operations electrodes and a magnet are arranged in a droplet actuator to manipulate a bead-containing droplet and position it relative to a magnetic field region that attracts the magnetically responsive beads. The droplet operations electrodes are operated to control the droplet shape and transport it away from the magnetic field region to form a concentration of beads in the droplet. The continued transport of the droplet away from the magnetic field causes the concentration of beads to break away from the droplet to yield a small, concentrated bead-containing droplet immobilized by the magnet.

Bead incubation and washing on a droplet actuator

The present invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay.

Droplet actuator with improved top substrate

The invention provides a droplet actuator comprising a base substrate and a top substrate separated to form a gap, wherein: (a) the base substrate comprises electrodes configured for conducting droplet operations in the gap; and (b) the top substrate comprises a first portion coupled to second portion, where the second portion comprises one or more openings establishing a fluid path extending from an exterior of the droplet actuator and into the gap. The invention also provides related methods of manufacturing the droplet actuator, methods of using the droplet actuator, and methods of loading the droplet actuator.

Droplet actuator configurations and methods of conducting droplet operations

Microfluidic feedback using impedance detection

Method of washing beads

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include providing a droplet including magnetically responsive beads. The droplet may be provided within a region of a magnetic field having sufficient strength to attract the magnetically responsive beads to an edge of the droplet or towards an edge of the droplet, or otherwise regionalize or aggregate beads within the droplet. The method may also include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads.

Droplet actuators, modified fluids and methods

The present invention provides droplet actuators, modified fluids and methods relating to droplet operations. An aspect includes a droplet actuator including a droplet operations substrate; an oil based filler fluid on the droplet operations substrate comprising an oil soluble additive in the filler fluid; and a droplet in contact with the oil based filler fluid. Still other aspects are provided.

Droplet actuator and droplet-based techniques

The invention is directed to certain droplet actuated molecular techniques. In one embodiment, the invention provides droplet actuator methods for detection of single nucleotide polymorphisms (SNPs) in a DNA sequence using digital microfluidics, including droplet actuator-based sample preparation and SNP analysis. In another embodiment, the invention provides droplet actuator devices and methods for providing integrated sample preparation and multiplexed detection of an infectious agent, such as HIV. In yet another embodiment, the invention provides droplet actuator devices and techniques for PCR amplification and detection of specific nucleic acid sequences using digital microfluidics, including droplet actuator-based sample preparation and target nucleic acid analysis. In yet another embodiment the invention provides methods for performing hot-start PCR on a droplet actuator. In yet another embodiment, the method of the invention combines PCR amplification with pyrosequencing to investigate specific sequences.

Method of making nucleic acid libraries

A method of preparing a nucleic acid library in droplets in contact with oil, including: (a) blunt-ending nucleic acid fragments in a droplet in the oil to yield blunt-ended nucleic acid fragments; (b) phosphorylating the blunt-ended nucleic acid fragments in a droplet in the oil to yield phosphorylated nucleic acid fragments; coupling A-tails to the phosphorylated nucleic acid fragments in a droplet in the oil to yield A-tailed nucleic acid fragments; and (d) coupling nucleic acid adapters to the A-tailed nucleic acid fragments in a droplet in the oil to yield the nucleic acid library comprising adapter-ligated nucleic acid fragments.

Microfluidic feedback using impedance detection

Methods comprising measuring the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode are disclosed. Computer readable mediums storing processor executable instructions for performing the method, and systems are also disclosed. The systems comprise a processor, memory and code stored in the memory that when executed cause the processor at least to: receive an output voltage signal, superimpose an excitation signal onto the output voltage signal to produce a superimposed signal, connect the superimposed signal to an electrode in a droplet actuator, suppress the output voltage signal, when detecting an impedance of the electrode, and measure the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode.

DROPLET AUTHOR SETTINGS AND METHODS FOR CONDUCTING DROPLET OPERATIONS.

CONFIGURAÇÕES DE ATUADOR DE GOTÍCULAS E MÉTODOS PARA CONDUZIR OPERAÇÕES DE GOTÍCULA. Trata-se de um atuador de gotícula com uma configuração de eletrodo de formação de gotícula associado a uma superfície de operações de gotícula, onde a configuração de eletrodo compreende um ou mais eletrodos configurados para controlar o volume de uma gotíclua durante a formação de uma sub-gotícula sobre a superfície de operações de gotícula. Métodos de fabricação e utilização do atuador de gotícula também são proporcionados. DROPOLE ACTUATOR SETTINGS AND METHODS FOR CONDUCTING DROPOLE OPERATIONS. It is a droplet actuator with a droplet forming electrode configuration associated with a droplet operations surface, where the electrode configuration comprises one or more electrodes configured to control the volume of a droplet during formation of a sub -droplet on the surface of droplet operations. Methods of manufacturing and using the droplet actuator are also provided.

Method of making nucleic acid libraries

A method of preparing a nucleic acid library in droplets in contact with oil, including: (a) blunt-ending nucleic acid fragments in a droplet in the oil to yield blunt-ended nucleic acid fragments; (b) phosphorylating the blunt-ended nucleic acid fragments in a droplet in the oil to yield phosphorylated nucleic acid fragments; coupling A-tails to the phosphorylated nucleic acid fragments in a droplet in the oil to yield A-tailed nucleic acid fragments; and (d) coupling nucleic acid adapters to the A-tailed nucleic acid fragments in a droplet in the oil to yield the nucleic acid library comprising adapter-ligated nucleic acid fragments.

Digital microfluidic device, system and method for performing a plasmonic particle-assisted elisa self-test

The present disclosure provides a digital microfluidic (DMF) cartridge for performing a self-test for a target analyte, including a DMF cartridge comprising a bottom substrate and a top substrate separated by a droplet operations gap, wherein the bottom substrate comprises a plurality of droplet operations electrodes configured for performing droplet operations on a liquid droplet in the droplet operations gap; one or more reaction chambers or reaction zones on the bottom substrate that are supplied by an arrangement of the droplet operations electrodes, wherein each reaction chamber or reaction zone comprises at least one detection spot and is configured for performing a plasmonic particle-assisted ELISA (pELISA) for detection and quantification of a target analyte in a sample droplet. The device may include downloadable software for a self-test and be operable using a smart device.

Droplet actuator devices and methods for manipulating beads

The invention provides droplet actuators and droplet actuator techniques. Among other things, the droplet actuators and methods are useful for manipulating beads on a droplet actuator, such as conducting droplet operations using bead-containing droplets on a droplet actuator. For example, beads may be manipulated on a droplet actuator in the context of executing a sample preparation protocol and/or an assay protocol. An output of the methods of the invention may be beads prepared for execution of an assay protocol. Another output of the methods of the invention may be results of an assay protocol executed using beads. Among the methods described herein are methods of concentrating beads in droplets, methods of washing beads, methods of suspending beads, methods of separating beads, methods of localizing beads within a droplet, methods of forming emulsions in which droplets include beads, methods of loading beads into a droplet operations gap of a droplet actuator, methods of organizing beads in a monolayer, and methods of capturing, trapping or restraining beads.

Bead manipulation techniques

The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include providing a droplet including magnetically responsive beads. The droplet may be provided within a region of a magnetic field having sufficient strength to attract the magnetically responsive beads to an edge of the droplet or towards an edge of the droplet, or otherwise regionalize or aggregate beads within the droplet. The method may also include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode -mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads.

Bubble Techniques for a Droplet Actuator

The present invention is directed to a droplet actuator and methods of making and using the droplet actuator including one or more substrates configured to form a droplet operations gap and including a physical or chemical feature that may be provided at a predetermined locus within or exposed to the droplet operations gap and configured to retain a bubble in position within the droplet operations gap.

Digital microfluidic device, system and method for performing a plasmonic particle-assisted elisa self-test

The present disclosure provides a digital microfluidic (DMF) cartridge for performing a self-test for a target analyte, including a DMF cartridge comprising a bottom substrate and a top substrate separated by a droplet operations gap, wherein the bottom substrate comprises a plurality of droplet operations electrodes configured for performing droplet operations on a liquid droplet in the droplet operations gap; one or more reaction chambers or reaction zones on the bottom substrate that are supplied by an arrangement of the droplet operations electrodes, wherein each reaction chamber or reaction zone comprises at least one detection spot and is configured for performing a plasmonic particle-assisted ELISA (pELISA) for detection and quantification of a target analyte in a sample droplet. The device may include downloadable software for a self-test and be operable using a smart device.

Droplet actuator configurations and methods of conducting droplet operations

A droplet actuator with a droplet formation electrode configuration associated with a droplet operations surface, wherein the electrode configuration comprises one or more electrodes configured to control volume of a droplet during formation of a sub-droplet on the droplet operations surface. Methods of making and using the droplet actuator are also provided.

Method of making nucleic acid libraries

Droplet actuator devices and methods

The invention provides droplet actuator devices for facilitating certain droplet actuated molecular techniques. In one embodiment, the invention provides droplet actuators that facilitate droplet operations in three dimensions. In another embodiment, the invention provides for cost-effective production of droplet actuators. In another embodiment, the invention provides for replacing one or more components of a droplet actuator. In another embodiment, the invention provides droplet actuators using printed conductive inks to form electrodes and/or ground planes. In another embodiment, the invention provides a magnetic clamping fixture for assembling droplet actuators. In another embodiment, the invention provides simple, low cost power sources for use in combination with microfluidic systems. In another embodiment, the invention provides an immunoassay multiplexing platform that uses digital microfluidics and real-time imaging of flash-based chemiluminescent signals. In still another embodiment, the invention provides droplet actuator devices that are fabricated using a plasma treatment process to raise substrate surface energy.

Bead incubation and washing on a droplet actuator

Methods are provided for separating magnetically responsive beads from a droplet in a droplet actuator. Droplet operations electrodes and a magnet are arranged in a droplet actuator to manipulate a bead-containing droplet and position it relative to a magnetic field region that attracts the magnetically responsive beads. The droplet operations electrodes are operated to control the droplet shape and transport it away from the magnetic field region to form a concentration of beads in the droplet. The continued transport of the droplet away from the magnetic field causes the concentration of beads to break away from the droplet to yield a small, concentrated bead-containing droplet immobilized by the magnet.

Method of making nucleic acid libraries

A method of preparing a nucleic acid library in droplets in contact with oil, including: (a) blunt-ending nucleic acid fragments in a droplet in the oil to yield blunt-ended nucleic acid fragments; (b) phosphorylating the blunt-ended nucleic acid fragments in a droplet in the oil to yield phosphorylated nucleic acid fragments; coupling A-tails to the phosphorylated nucleic acid fragments in a droplet in the oil to yield A-tailed nucleic acid fragments; and (d) coupling nucleic acid adapters to the A-tailed nucleic acid fragments in a droplet in the oil to yield the nucleic acid library comprising adapter-ligated nucleic acid fragments.

Droplet Actuators, Modified Fluids and Methods

The present invention provides droplet actuators, modified fluids and methods relating to droplet operations. An aspect includes a droplet actuator including a droplet operations substrate; an oil based filler fluid on the droplet operations substrate comprising an oil soluble additive in the filler fluid; and a droplet in contact with the oil based filler fluid. Still other aspects are provided.

Droplet Actuator Configurations and Methods of Conducting Droplet Operations

A droplet actuator with a droplet formation electrode configuration associated with a droplet operations surface, wherein the electrode configuration comprises one or more 5 electrodes configured to control volume of a droplet during formation of a sub-droplet on the droplet operations surface. Methods of making and using the droplet actuator are also provided. 210a 214 218a 210b As- -Figure 2A 234 218b 222 210a 214 218a 210b _ ___ - Figue_2 234 218b 222 210a214 18a210b 230a FF 230b Axis ure -- - FF- - Figure 2C 234 218b 222

Droplet actuator configurations and methods of conducting droplet operations.

Un aplicador de gotas con una configuración de electrodo de formación de gotas asociado con una superficie de operaciones de gotas, en donde las configuraciones de electrodo comprende uno o mas electrodos configurados para controlar el volumen de una gota durante la formación de una sub-gota en la superficie de operaciones de gotas. También se proporcionan métodos para hacer y usar el aplicador de gotas.

Plasmon resonance (pr) system and instrument, digital microfluidic (dmf) cartridge, and methods of using localized surface plasmon resonance (lspr) for analysis of analytes

A plasmon resonance (PR) system and instrument, digital microfluidic (DMF) cartridge, and methods of using localized surface plasmon resonance (LSPR) and droplet operations for analysis of analytes is disclosed. For example, a PR system is provided that may include a DMF cartridge that may support both fixed LSPR sensing capability and in-solution LSPR sensing capability for analysis of analytes. The DMF cartridge may include an electrode arrangement for performing droplet operations, wherein the droplet operations can be used for performing fixed LSPR sensing operations and in-solution LSPR sensing operations. Further, methods of using droplet operations in the DMF cartridge to perform fixed LSPR sensing operations and/or in-solution LSPR sensing operations are provided.

Colored coverlay dielectrics in digital microfluidics

Provided herein are devices, systems and methods comprising a digital microfluidics (DMF) cartridge, the DMF cartridge comprising: a top plate, wherein the top plate comprises a top plate substrate; a bottom plate, wherein the bottom plate comprises a colored coverlay substrate; and a droplet operation gap between the top plate and the bottom plate.

Pipette dispenser system and method

A pipette dispenser vision system and method are disclosed. For example, a pipette dispenser vision system is provided for tracking pipettes with respect to dispensing liquid into target fluid wells, vessels, and/or reservoirs. The presently disclosed pipette dispenser vision system may include, for example, a computing device, a red/green/blue (RGB) imaging device, an infrared (IR) imaging device, an IR illumination source, and an IR sensor; all arranged with respect to, for example, a single-channel and/or a multi-channel pipette for dispensing liquid into a dispensing platform. Further, a method of using the presently disclosed pipette dispenser vision system is provided. For example, the method processes image data from the RGB imaging device and/or the IR imaging device to monitor/verify certain operations of the pipetting process and/or dispensing platform.

Microfluidics device and method including bottom substrate, top substrate, and cover plate

Provided herein are devices comprising: a bottom plate comprising one or more electrodes configured for performing droplet operations in a droplet operations gap formed between the bottom plate and a top plate, the top plate having a top surface comprising one or more irregularities or protrusions such the top surface is non-planar; and a cover plate configured to be disposed over the top plate, the cover plate comprising one or more alignment features for positioning the cover plate with respect to the one or more irregularities or protrusions of the top surface of the top plate.

Plasmon resonance (pr) system and instrument, digital microfluidic (dmf) cartridge, and methods of using localized surface plasmon resonance (lspr) for analysis of analytes

A plasmon resonance (PR) system and instrument, digital microfluidic (DMF) cal lt idge, and methods of using localized surface plasmon resonance (LSPR) and droplet operations for analysis of analytes is disclosed. For example, a PR system is provided that may include a DMF cal tiidge that may support both fixed LSPR sensing capability and in-solution LSPR sensing capability for analysis of analytes. The DMF cartridge may include an electrode arrangement for performing droplet operations, wherein the droplet operations can be used for performing fixed LSPR sensing operations and in-solution LSPR sensing operations. Further, methods of using droplet operations in the DMF cal tiidge to perform fixed LSPR sensing operations and/or in-solution LSPR sensing operations are provided.

Digital microfluidic device, system and method for performing a plasmonic particle-assisted elisa self-test

Digital microfluidics cartridge, system, and method

A cartridge with (a) a top substrate and a bottom substrate separated to form a droplet operations gap, the top substrate comprising a well plate comprising an array of wells each well comprising a hollow needle liquidly connecting an interior of the well with the droplet operations gap; and (b) a compressible membrane layer atop the well plate sealing the reservoirs and arranged so that compression of the flexible membrane towards the interior of the well forces a liquid from the interior of the well through the hollow needle, and into the droplet operations gap.

Pipette dispenser system and method

A pipette dispenser vision system and method are disclosed. For example, a pipette dispenser vision system is provided for tracking pipettes with respect to dispensing liquid into target fluid wells, vessels, and/or reservoirs. The presently disclosed pipette dispenser vision system may include, for example, a computing device, a red/green/blue (RGB) imaging device, an infrared (IR) imaging device, an IR illumination source, and an IR sensor; all arranged with respect to, for example, a single-channel and/or a multi-channel pipette for dispensing liquid into a dispensing platform. Further, a method of using the presently disclosed pipette dispenser vision system is provided. For example, the method processes image data from the RGB imaging device and/or the IR imaging device to monitor/verify certain operations of the pipetting process and/or dispensing platform.

Pipette dispenser system and method

A pipette dispenser vision system and method are disclosed. For example, a pipette dispenser vision system is provided for tracking pipettes with respect to dispensing liquid into target fluid wells, vessels, and/or reservoirs. The presently disclosed pipette dispenser vision system may include, for example, a computing device, a red/green/blue (RGB) imaging device, an infrared (IR) imaging device, an IR illumination source, and an IR sensor; all arranged with respect to, for example, a single-channel and/or a multi-channel pipette for dispensing liquid into a dispensing platform. Further, a method of using the presently disclosed pipette dispenser vision system is provided. For example, the method processes image data from the RGB imaging device and/or the IR imaging device to monitor/verify certain operations of the pipetting process and/or dispensing platform.

Ligand assisted dissociation

Provided herein are methods, systems and digital microfluidic cartridges for the determination of ligand-target analyte affinity value (KD), determination of an association rate constant (kon), and/or determination of a dissociation rate constant (koff) in a PR (plasmon resonance) based system. More specifically, as described herein, the present disclosure is directed to improved methods, systems and digital microfluidic cartridges for determining a dissociation rate constant (koff) using free ligand molecules in the dissociation phase or dissociation step of a standard kinetic binding curve or in ligand-target analyte affinity analysis (referred to herein as ligand assisted dissociation (LAD)).

Pipette dispenser system and method

A pipette dispenser vision system and method are disclosed. For example, a pipette dispenser vision system is provided for tracking pipettes with respect to dispensing liquid into target fluid wells, vessels, and/or reservoirs. The presently disclosed pipette dispenser vision system may include, for example, a computing device, a red/green/blue (RGB) imaging device, an infrared (IR) imaging device, an IR illumination source, and an IR sensor; all arranged with respect to, for example, a single-channel and/or a multi-channel pipette for dispensing liquid into a dispensing platform. Further, a method of using the presently disclosed pipette dispenser vision system is provided. For example, the method processes image data from the RGB imaging device and/or the IR imaging device to monitor/verify certain operations of the pipetting process and/or dispensing platform.

Pipette dispenser system and method

A pipette dispenser vision system and method are disclosed. For example, a pipette dispenser vision system is provided for tracking pipettes with respect to dispensing liquid into target fluid wells, vessels, and/or reservoirs. The presently disclosed pipette dispenser vision system may include, for example, a computing device, a red/green/blue (RGB) imaging device, an infrared (LR) imaging device, an IR illumination source, and an IR sensor; all arranged with respect to, for example, a single-channel and/or a multi-channel pipette for dispensing liquid into a dispensing platform. Further, a method of using the presently disclosed pipette dispenser vision system is provided. For example, the method processes image data from the RGB imaging device and/or the IR imaging device to monitor/verify certain operations of the pipetting process and/or dispensing platform.

Pipette dispenser system and method

A pipette dispenser vision system and method are disclosed. For example, a pipette dispenser vision system is provided for tracking pipettes with respect to dispensing liquid into target fluid wells, vessels, and/or reservoirs. The presently disclosed pipette dispenser vision system may include, for example, a computing device, a red/green/blue (RGB) imaging device, an infrared (IR) imaging device, an IR illumination source, and an IR sensor; all arranged with respect to, for example, a single-channel and/or a multi-channel pipette for dispensing liquid into a dispensing platform. Further, a method of using the presently disclosed pipette dispenser vision system is provided. For example, the method processes image data from the RGB imaging device and/or the IR imaging device to monitor/verify certain operations of the pipetting process and/or dispensing platform.

Droplet actuator configurations and methods

Droplet actuators for conducting droplet operations, such as droplet transport and droplet dispensing, are provided. In one embodiment, the droplet actuator may include a substrate including, droplet operations electrodes arranged for conducting droplet operations on a surface of the substrate; and reference electrodes associated with the droplet operations electrodes and extending above the surface of the substrate. Other embodiments of droplet actuators and methods of loading and using such droplet actuators are also provided.

Optical analysis on digital microfluidic (dmf) cartridges

Provided herein are digital microfluidic devices, methods and systems for improving absorbance and/or transmission detection in electromagnetic radiation spectroscopy. For example, devices and methods are provided for determining the absorbance and/or transmission of light when analyzing a fluid (e.g., a droplet) including a target analyte of interest.

Pipette dispenser system and method

Analysis cartridge cover plate

Analysis cartridge top plate

Analysis cartridge cover plate