Microfluidic free interface diffusion techniques

A static fluid and a second fluid are placed into contact along a microfluidic free interface and allowed to mix by diffusion without convective flow across the interface. In accordance with one embodiment of the present invention, the fluids are static and initially positioned on either side of a closed valve structure in a microfluidic channel having a width that is tightly constrained in at least one dimension. The valve is then opened, and no-slip layers at the sides of the microfluidic channel suppress convective mixing between the two fluids along the resulting interface. Applications for microfluidic free interfaces in accordance with embodiments of the present invention include, but are not limited to, protein crystallization studies, protein solubility studies, determination of properties of fluidics systems, and a variety of biological assays such as diffusive immunoassays, substrate turnover assays, and competitive binding assays.

High throughput screening of crystallization of materials

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels.

MICROFABRICATED ELASTOMERIC VALVE AND PUMP SYSTEMS

A method of fabricating an elastomeric structure, comprising: forming a first elastomeric layer on top of a first micromachined mold, the first micromachined mold having a first raised protrusion which forms a first recess extending along a bottom surface of the first elastomeric layer; forming a second elastomeric layer on top of a second micromachined mold, the second micromachined mold having a second raised protrusion which forms a second recess extending along a bottom surface of the second elastomeric layer; bonding the bottom surface of the second elastomeric layer onto a top surface of the first elastomeric layer such that a control channel forms in the second recess between the first and second elastomeric layers; and positioning the first elastomeric layer on top of a planar substrate such that a flow channel forms in the first recess between the first elastomeric layer and the planar substrate. 1. (canceled)2. The method of using a microfluidic device , comprising:flowing a sample into a looped channel of the microfluidic device;pumping the sample around the loop channel with a peristaltic action, wherein the peristaltic action comprises sequential activation of a series of individually addressable valves arranged along the looped channel.3. The method of claim 2 , wherein the series of individually addressable valves comprise three valves.4. The method of claim 3 , wherein each of the three valves is an intersection of an individually addressable control line over the looped channel.5. The method of claim 2 , wherein the microfluidic device is an elastomeric device.6. The method of claim 2 , wherein the pumping passes the sample over a DNA array. This nonprovisional patent application is a continuation of U.S. application Ser. No. 14/188,664 filed Feb. 24, 2014, which is a continuation of Ser. No. 11/932,263 filed Oct. 31, 2007, now U.S. Pat. No. 8,656,958, which is a continuation of U.S. application Ser. No. 11/685,654 filed Mar. 13, 2007, now U.S. Pat. ...

SYSTEM AND METHOD FOR MICROFLUIDIC CELL CULTURE

Microfluidic devices and methods for perfusing a cell with perfusion fluid are provided herein, wherein the gravitational forces acting on the cell to keep the cell at or near a retainer or a retaining position exceed the hydrodynamic forces acting on the cell to move it toward an outlet. 2. The method of claim 1 , wherein the speed of the perfusion fluid is decreased to less than 50 μm/s as the perfusion fluid approaches the retaining position.3. The method of claim 1 , wherein the speed of the perfusion fluid is decreased to about 0 μm/s as the perfusion fluid approaches the retaining position.4. The method of claim 1 , wherein the chamber has a top and a bottom claim 1 , and the retaining position is at the bottom.5. The method of claim 4 , wherein the inlet position is proximal to the top.6. The method of or claim 4 , wherein the outlet position is proximal to the top.7. The method of claim 4 , wherein the cell is retained at the bottom by gravitational forces.8. The method of claim 1 , further comprising regulating osmolarity of the perfusion fluid within the chamber.9. The method of claim 8 , wherein regulating osmolarity of the perfusion fluid within the chamber comprises placing the chamber in fluid communication with a bathing fluid claim 8 , wherein the bathing fluid has a volume greater than the chamber volume.10. The method of claim 9 , wherein the bathing fluid and the perfusion fluid are iso-osmotic.11. The method of claim 1 , wherein the cell is a suspension cell.12. The method of claim 1 , wherein the perfusion fluid comprises any one or more of a cell culture medium claim 1 , an immunostaining agent claim 1 , an enzymatic reagent claim 1 , a dye claim 1 , an oil claim 1 , and a bead-containing solution.13. The method of claim 1 , wherein a length of the first region is less than or equal to a length of the shortest distance between the retaining position and the first region.14. The method of claim 13 , wherein the ratio of the length of the first ...

MICROFLUIDIC SIEVE VALVES

Sieve valves for use in microfluidic device are provided. The valves are useful for impeding the flow of particles, such as chromatography beads or cells, in a microfluidic channel while allowing liquid solution to pass through the valve. The valves find particular use in making microfluidic chromatography modules. 16-. (canceled)7. A microfluidic device comprising two or more sieve valves , wherein a chromatographic separation medium is disposed between two sieve valves thereby forming a separation column.8. (canceled)9. The device of that comprises more than 20 separation columns.10. A microfluidic device comprising a microfluidic chromatography column claim 7 , said column comprising a chromatographic separation medium disposed behind a sieve valve claim 7 , and optionally disposed between two sieve valves.11. The device of wherein the chromatographic separation medium comprises a polymeric bead coupled to a ligand.12. The device of wherein the beads have been derivatized to bind a nucleic acid.13. The device of wherein the beads have been derivatized with oligo(dT).14. The device of wherein the beads have been derivatized with a protein claim 10 , optionally an antibody.15. The device of that contains five or more sieve valves paired with conventional valves.16. A microfluidic device comprising two or more sieve valves paired with conventional valves.1720-. (canceled) This application claims benefit of U.S. Provisional Application No. 60/633,121, filed Dec. 3, 2004, the entire contents of which are incorporated herein by reference.Work described herein has been supported, in part, by National Institutes of Health (NIH) grant NIH 1RO1 HG002644-01A1. The United States government may have certain rights in the invention.The present invention relates to microfabricated devices and microchromatography.Microfluidic devices may be used in a variety of biomedical and pharmaceutical applications, including analysis, preparation and synthesis of chemical compounds and ...

Microfluidic Protein Crystallography Techniques

The present invention relates to microfluidic devices and methods facilitating the growth and analysis of crystallized materials such as proteins. In accordance with one embodiment, a crystal growth architecture is separated by a permeable membrane from an adjacent well having a much larger volume. The well may be configured to contain a fluid having an identity and concentration similar to the solvent and crystallizing agent employed in crystal growth, with diffusion across the membrane stabilizing that process. Alternatively, the well may be configured to contain a fluid having an identity calculated to affect the crystallization process. In accordance with the still other embodiment, the well may be configured to contain a material such as a cryo-protectant, which is useful in protecting the crystalline material once formed.

Multiplex Amplification for the Detection of Nucleic Acid Variations

Kits, primers, and methods are provided herein for detecting relative target source to reference source ratios in a biological sample, by distributing the biological sample into discrete subsamples, wherein the biological sample includes, a plurality of target molecules on a target source; and a plurality of reference molecules on a reference source; providing target primers directed to one or more of the plurality of target molecules and reference primers directed to one or more of the plurality of reference molecules; performing digital amplification with the target primers and the reference primers; and detecting the presence or absence of amplified target products with target probes and detecting the presence or absence of amplified reference products with reference probes, wherein the ratio of amplified target products to amplified reference products is indicative of a relative amount of target source to reference source in a biological sample. 1. A method of detecting relative target source to reference source ratios in a biological sample , the method comprising: a plurality of target molecules corresponding to spaced apart target sites on a target source; and', 'a plurality of reference molecules corresponding to spaced apart reference sites on a reference source;, '(a) distributing the biological sample into discrete subsamples, wherein the biological sample comprises'}(b) providing a target primer subset of at least one digital amplification target primer pair directed to one or more of the plurality of target molecules and a reference primer subset of at least one digital amplification reference primer pair directed to one or more of the plurality of reference molecules;(c) performing digital amplification with the target primer subset and the reference primer subset; and(d) detecting the presence or absence of amplified target products with a subset of target probes and detecting the presence or absence of amplified reference products with a subset of ...

SYSTEM AND METHOD FOR MICROFLUIDIC CELL CULTURE

Microfluidic devices and methods for perfusing a cell with perfusion fluid are provided herein, wherein the gravitational forces acting on the cell to keep the cell at or near a retainer or a retaining position exceed the hydrodynamic forces acting on the cell to move it toward an outlet. Also provided, are methods for assaying cell products within the microfluidic device. 1. A method , the method comprising:(a) retaining a cell at a retaining position within a microfluidic chamber having a chamber volume;(b) flowing a perfusing fluid through the microfluidic chamber, wherein the perfusing fluid enters the chamber through an inlet at an inlet position and exits the chamber through an outlet at an outlet position; and(c) measuring a cell product produced by the cell within the microfluidic chamber;wherein the perfusing fluid has a greater velocity laminar flow adjacent the inlet and outlet positions than at the retaining position, and wherein a first region of the chamber is spaced apart from the retaining position, wherein the first region is interposed directly between the inlet and outlet positions.2. The method of claim 1 , wherein measuring the cell product may be selected from one or more of: lineage staining; cell-surface protein staining; antibody staining; enzymatic assay; RT-PCR analysis; PCR analysis; sequencing; functional assay; and bead capture assay to characterize the cells.3. A method claim 1 , the method comprising:(a) retaining a cell at a retaining position within a microfluidic chamber;(b) flowing a perfusion fluid into the microfluidic chamber through an inlet;(c) flowing the perfusion fluid out of the microfluidic chamber through an outlet wherein the outlet is positioned such that gravitational forces acting on the cell to keep it at or near the retaining position exceed hydrodynamic forces acting on the cell to move it toward the outlet; and(d) measuring a cell product produced by the cell within the microfluidic chamber.4. The method of ...

HIGH THROUGHPUT SCREENING OF CRYSTALLIZATION OF MATERIALS

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels. 1loading a plurality of wells on an upper surface of a microfluidic device with a liquid material;biasing a holder piece against the upper surface such that a continuous raised rim of the holder piece presses against the upper surface surrounding the wells, such that a chamber is created over the wells; andapplying a positive pressure to the chamber to drive the material from the wells into an active area of the elastomeric microfluidic structure.. A method of priming a microfluidic device with a liquid material, the method comprising: This application is a continuation of U.S. patent application Ser. No. 11/668,263, filed Jan. 29, 2007, (now U.S. Pat. No. 8,382,896), which is a divisional of U.S. patent application Ser. No. 10/117,978 filed Apr. 5, 2002, (now U.S. Pat. No. 7,195,670), which claims the benefit of U.S. Provisional Patent Application No. 60/323,524, filed Sep. 17, 2001, and which is a continuation-in-part of U.S. patent application Ser. No. 09/887,997 filed Jun. 22, 2001, (now U.S. Pat. No. 7,052,545), which is a continuation-in-part of U.S. patent application Ser. No. 09/826,583 filed Apr. 6, 2001, (now U.S. Pat. No. 6,899,137), ...

METHODS FOR ASSAYING CELLULAR BINDING INTERACTIONS

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 1. A method of assaying for a binding interaction between a protein produced by a cell and a biomolecule:(a) retaining the cell within a chamber having an inlet and an outlet;(b) exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell;(c) flowing a first fluid volume comprising the biomolecule through the inlet into the chamber and out the outlet, wherein the first fluid volume is in fluid communication with the capture substrate; and(d) determining binding interactions between the protein produced by a cell and the biomolecule.2. The method of claim 1 , wherein the cell is an antibody producing cell (APC) claim 1 , the protein produced by the cell is an antibody and the biomolecule is an antigen.3. The method of or claim 1 , wherein the biomolecule is a fluorescently labeled antigen.4. The method of claim 1 , wherein the determining binding interactions is a measure of antigen-antibody binding kinetics.5. The method of claim 4 , wherein the measure of antigen-antibody binding kinetics is any one or both of: a Krate; and a Krate.6 ...

METHODS FOR ASSAYING CELLULAR BINDING INTERACTIONS

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145.-. (canceled)46. A method of assaying for a binding interaction between an antibody secreted by a single antibody secreting cell (ASC) and an antigen , the method comprising:retaining the single ASC within a chamber having a volume of from 100 μL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single ASC within the chamber to produce a secreted antibody;bringing a first fluid volume comprising the antigen in fluid communication with the secreted antibody;exposing the secreted antibody to a removeable capture substrate, wherein the removeable capture substrate is in fluid communication with the secreted antibody and wherein the removeable capture substrate is operable to bind the secreted antibody;incubating the secreted antibody with the removeable capture substrate to produce a bound antibody; andmeasuring a binding interaction between the secreted antibody and the antigen.47. The method of claim 46 , wherein the single APC is a single primary plasma cell.48. The method of claim 46 , wherein the single APC is an activated B cell.49. The method of claim 46 , wherein the antigen is a cell or a cell fragment.50. The method of claim 46 , wherein the antigen is a virus or ...

MICROFLUIDIC CHEMOSTAT

A chemostat is described that includes a growth chamber having a plurality of compartments. Each of the compartments may be fluidly isolated from the rest of the growth chamber by one or more actuatable valves. The chemostat may also include a nutrient supply-line to supply growth medium to the growth chamber, and an output port to remove fluids from the growth chamber. Also, a method of preventing biofilm formation in a growth chamber of a chemostat is described. The method may include the steps of adding a lysis agent to a isolated portion of the growth chamber, and reuniting the isolated portion with the rest of the growth chamber. 1. A chemostat comprising:a growth chamber having a plurality of compartments, wherein each of the compartments may be fluidly isolated from the rest of the growth chamber by one or more actuatable valves;a nutrient supply-line to supply growth medium to the growth chamber; andan output port to remove fluids from the growth chamber.2. A chemostat chip comprising an array of chemostats , wherein each of the chemostats comprises:a growth chamber having a plurality of compartments, wherein each of the compartments may be fluidly isolated from the rest of the growth chamber by one or more actuatable valves;a nutrient supply-line to supply growth medium to the growth chamber; andan output port to remove fluids from the growth chamber.3. A method of preventing biofilm formation in a growth chamber of a chemostat , the method comprising:adding a lysis agent to an isolated portion of the growth chamber; andreuniting the isolated portion with the rest of the growth chamber. This application is a continuation of U.S. patent application Ser. No. 13/197,654, filed Aug. 3, 2011, titled “MICROFLUIDIC CHEMOSTAT” which is a continuation of U.S. patent application Ser. No. 12/182,088, filed Jul. 29, 2008, titled “MICROFLUIDIC CHEMOSTAT,” which is a divisional of U.S. patent application Ser. No. 11/012,852, filed Dec. 14, 2004, titled “MICROFLUIDIC ...

NUCLEIC ACID AMPLIFICATION USING MICROFLUIDIC DEVICES

The present invention provides microfluidic devices and methods using the same in various types of thermal cycling reactions. Certaom devices include a rotary microfluidic channel and a plurality of temperature regions at different locations along the rotary microfluidic channel at which temperature is regulated. Solution can be repeatedly passed through the temperature regions such that the solution is exposed to different temperatures. Other microfluidic devices include an array of reaction chambers formed by intersecting vertical and horizontal flow channels, with the ability to regulate temperature at the reaction chambers. The microfluidic devices can be used to conduct a number of different analyses, including various primer extension reactions and nucleic acid amplification reactions. 1. A microfluidic device , comprising:(a) a substrate comprising an elastomeric material; configured such that a sample introduced, into the flow channel can be cycled around the flow channel; and', 'comprising a plurality of temperature regions at which temperature can be regulated, each temperature region located at a different location along the flow channel;, '(b) a flow channel disposed within the substrate, the flow channel'}(c) an inlet in fluid communication with the flow channel via which the sample can be introduced into the flow channel; and(d) a temperature controller operatively disposed to regulate temperature within at least one of the plurality of temperature regions.299-. (canceled) This application claims the benefit of U.S. Provisional Application No. 60/281,960, filed Apr. 6, 2001, U.S. Provisional Application No. 60/300,516, filed Jun. 22, 2001, and U.S. Provisional Application No. 60/334,473, filed Nov. 16, 2001, each of which is incorporated herein by reference in its entirety for all purposes.This invention was made with support from Grant Number CTS-0088649 awarded by the National Science Foundation. Therefore, the U.S. government has certain rights in ...

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 1. A method of assaying for a binding interaction between a protein secreted by a cell and a biomolecule , the method comprising:(a) retaining the cell secreting the protein within a chamber having a solid wall defining the chamber;(b) exposing the protein secreted by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein secreted by the cell and wherein the capture substrate is operable to bind the protein secreted by the cell to produce a bound protein;(c) flowing a first fluid volume comprising the biomolecule into the chamber over the bound protein, wherein the first fluid volume is in fluid communication with the capture substrate and the bound protein; and(d) measuring a binding interaction between the protein produced by a cell and the biomolecule.2. The method of claim 1 , wherein the cell is an antibody producing cell (APC) claim 1 , the protein secreted by the cell is an antibody and the biomolecule is an antigen.3. The method of claim 1 , wherein the biomolecule is a fluorescently labeled antigen.4. The method of claim 1 , wherein measuring a binding interaction comprises measuring an antigen-antibody binding kinetic property.5. The method of claim 4 , ...

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145.-. (canceled)46. A method of assaying for a binding interaction between an antibody secreted by a single antibody secreting cell (ASC) and an antigen , the method comprising:retaining the single ASC within a chamber having a volume of less than 500 μL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single ASC within the chamber to produce a secreted antibody;bringing a first fluid volume comprising the antigen in fluid communication with the secreted antibody;exposing the secreted antibody to a removeable capture substrate, wherein the removeable capture substrate is in fluid communication with the secreted antibody and wherein the removeable capture substrate is operable to bind the secreted antibody;incubating the secreted antibody with the removeable capture substrate to produce a bound antibody; andmeasuring a binding interaction between the secreted antibody and the antigen.47. The method of claim 46 , wherein the single APC is a single primary plasma cell.48. The method of claim 46 , wherein the single APC is an activated B cell.49. The method of claim 46 , wherein the antigen is a cell or a cell fragment.50. The method of claim 46 , wherein the antigen is a virus or a ...

System and Method for Microfluidic Cell Culture

Microfluidic devices and methods for perfusing a cell with perfusion fluid are provided herein, wherein the gravitational forces acting on the cell to keep the cell at or near a retainer or a retaining position exceed the hydrodynamic forces acting on the cell to move it toward an outlet. Also provided, are methods for assaying cell products within the microfluidic device.

HIGH THROUGHPUT SCREENING OF CRYSTALLIZATION OF MATERIALS

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels. 16.-. (canceled)7. A microfluidic device comprising:a microfluidic flow channel;a control channel; andan elastomeric membrane moveable from a relaxed position blocking the flow channel to a actuated position in which the flow channel is open when the control channel is placed under a negative control pressure.8. The microfluidic device of claim 7 , wherein the elastomeric membrane comprises a separating portion which claim 7 , in a relaxed position claim 7 , separates a first portion of the flow channel from a second portion of the flow channel.9. The microfluidic device of claim 8 , wherein a portion of the control channel overlies the separating portion.10. The microfluidic device of claim 9 , wherein the separating portion is drawn into the control channel when the control channel is placed under the negative control pressure.11. The microfluidic device of claim 9 , wherein a first portion of the control channel overlies the separating portion and is wider than the separating portion.12. The microfluidic device of claim 11 , wherein the first portion of the control channel is wider than a second portion of the control channel that does not ...

METHODS FOR ASSAYING CELLULAR BINDING INTERACTIONS

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145.-. (canceled)46. A method of assaying for a binding interaction between an antibody produced by a single antibody producing cell (APC) and a biomolecule , the method comprising:retaining the single APC within a chamber having a volume of from 100 μL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;exposing the antibody produced by the APC to a capture substrate, wherein the capture substrate is in fluid communication with the antibody produced by the APC and wherein the capture substrate is operable to bind the antibody produced by the APC to produce a bound antibody;bringing a fluid volume comprising the biomolecule in fluid communication with the capture substrate and the bound antibody; andmeasuring a binding interaction between the antibody produced by the APC and the biomolecule.47. The method of claim 46 , wherein the single APC is a single primary B cell or a single memory B cell.48. The method of claim 46 , wherein the single APC is a single primary plasma cell.49. The method of claim 46 , wherein the single APC is from a human claim 46 , a rabbit claim 46 , a rat claim 46 , a mouse claim 46 , a sheep claim 46 , an ape claim 46 , a monkey claim 46 , a goat claim 46 , a dog ...

System And Method For Microfluidic Cell Culture

Microfluidic devices and methods for perfusing a cell with perfusion fluid are provided herein, wherein the gravitational forces acting on the cell to keep the cell at or near a retainer or a retaining position exceed the hydrodynamic forces acting on the cell to move it toward an outlet. Also provided, are methods for assaying cell products within the microfluidic device.

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145-. (canceled)46. A method of assaying for a binding interaction between a secreted monoclonal antibody produced by a single antibody producing cell (APC) and an antigen , the method comprising:retaining the single APC within a chamber having a volume of from 100 μL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single APC within the chamber to produce a secreted monoclonal antibody;exposing the secreted monoclonal antibody to a first removeable capture substrate bound to an antigen, wherein the antigen is capable of capturing the secreted monoclonal antibody,incubating the secreted monoclonal antibody with the first removeable capture substrate to produce a bound antibody; andmeasuring a binding interaction between the bound antibody and the antigen.47. The method of claim 46 , wherein the single APC is a primary B cell or a memory B cell.48. The method of claim 46 , wherein the single APC is a primary plasma cell.49. The method of claim 46 , wherein the single APC is from a human claim 46 , a rabbit claim 46 , a rat claim 46 , a mouse claim 46 , a sheep claim 46 , an ape claim 46 , a monkey claim 46 , a goat claim 46 , a dog claim 46 , a cat claim 46 , a camel claim ...

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule.

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145.-. (canceled)46. A method of assaying for a binding interaction between a secreted monoclonal antibody produced by a single antibody producing cell (APC) and an antigen , the method comprising:retaining the single APC within a chamber having a volume of from 100 pL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single APC within the chamber to produce a secreted monoclonal antibody;exposing the secreted monoclonal antibody to a first removeable capture substrate bound to an antigen, wherein the antigen is capable of capturing the secreted monoclonal antibody, incubating the secreted monoclonal antibody with the first removeable capture substrate to produce a bound antibody;measuring a binding interaction between the bound antibody and the antigen;lysing the single APC and capturing the nucleic acids of the single APC on a second removeable capture substrate.47. The method of claim 46 , wherein the single APC is a primary B cell or a memory B cell.48. The method of claim 46 , wherein the single APC is a primary plasma cell.49. The method of claim 46 , wherein the single APC is from a human claim 46 , a rabbit claim 46 , a rat claim 46 , a mouse claim 46 , a sheep ...

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145.-. (canceled)46. A method of assaying for a binding interaction between a secreted monoclonal antibody produced by a single antibody producing cell (APC) and an antigen , the method comprising:retaining the single APC within a chamber having a volume of from 100 μL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single APC within the chamber to produce a secreted monoclonal antibody;exposing the secreted monoclonal antibody to a first removeable capture substrate, wherein the first removeable capture substrate is in fluid communication with the secreted monoclonal antibody and wherein the first removeable capture substrate is capable of binding the secreted monoclonal antibody;incubating the secreted monoclonal antibody with the removeable capture substrate to produce a bound antibody;bringing a first fluid volume comprising the antigen in fluid communication with the bound antibody;measuring a binding interaction between the bound antibody and the antigen; andlysing the single APC and capturing the nucleic acids of the single APC on a second removeable capture substrate.47. The method of claim 46 , wherein the single APC is a primary B cell or a memory B cell.48. The ...

Microfluidic Devices and Methods for Use Thereof in Multicellular Assays of Secretion

Methods and devices are provided herein for identifying a cell population comprising an effector cell that exerts an extracellular effect. In one embodiment the method comprises retaining in a microreactor a cell population comprising one or more effector cells, wherein the contents of the microreactor further comprise a readout particle population comprising one or more readout particles, incubating the cell population and the readout particle population within the microreactor, assaying the cell population for the presence of the extracellular effect, wherein the readout particle population or subpopulation thereof provides a direct or indirect readout of the extracellular effect, and determining, based on the results of the assaying step, whether one or more effector cells within the cell population exerts the extracellular effect on the readout particle. If an extracellular effect is measured, the cell population is recovered for further analysis to determine the cell or cells responsible for the effect. 1218.-. (canceled)219. A method of identifying a cell population comprising an effector cell having an extracellular effect , comprising:retaining in a microreactor a cell population comprising one or more effector cells, wherein the contents of the microreactor further comprise a readout particle population comprising one or more readout particles,incubating the cell population and the one or more readout particles within the microreactor,assaying the cell population for the presence of the extracellular effect, wherein the readout particle population or a subpopulation thereof provides a direct or indirect readout of the extracellular effect, anddetermining, based on the results of the assaying step, whether one or more effector cells within the cell population exhibits the extracellular effect.220. The method of claim 219 , wherein the one or more effector cells comprise an antibody secreting cell (ASC).221. The method of claim 219 , wherein the cell ...

MICROFLUIDIC FREE INTERFACE DIFFUSION TECHNIQUES

A static fluid and a second fluid are placed into contact along a microfluidic free interface and allowed to mix by diffusion without convective flow across the interface. In accordance with one embodiment of the present invention, the fluids are static and initially positioned on either side of a closed valve structure in a microfluidic channel having a width that is tightly constrained in at least one dimension. The valve is then opened, and no-slip layers at the sides of the microfluidic channel suppress convective mixing between the two fluids along the resulting interface. Applications for microfluidic free interfaces in accordance with embodiments of the present invention include, but are not limited to, protein crystallization studies, protein solubility studies, determination of properties of fluidics systems, and a variety of biological assays such as diffusive immunoassays, substrate turnover assays, and competitive binding assays. 141-. (canceled)42. A method of creating a concentration gradient of a chemical species comprising:disposing a first fluid containing the chemical species;disposing a second static fluid proximate to the first static fluid to form a microfluidic free interface;suppressing convective flow of the first and second fluids such that mixing between the first and second fluids across the microfluidic free interface occurs substantially exclusively by diffusion and a concentration gradient of the chemical species is created.43. The method of wherein the first chemical species comprises a nutrient for a cell in fluidic communication with the second fluid.44. The method of further comprising:disposing a third fluid containing a second chemical species proximate to the second fluid to form a second microfluidic free interface;suppressing convective flow of the second and third fluids such that mixing between the second and third fluids across the second microfluidic free interface occurs substantially exclusively by diffusion and a second ...

MICROFABRICATED ELASTOMERIC VALVE AND PUMP SYSTEMS

A method of fabricating an elastomeric structure, comprising: forming a first elastomeric layer on top of a first micromachined mold, the first micromachined mold having a first raised protrusion which forms a first recess extending along a bottom surface of the first elastomeric layer; forming a second elastomeric layer on top of a second micromachined mold, the second micromachined mold having a second raised protrusion which forms a second recess extending along a bottom surface of the second elastomeric layer; bonding the bottom surface of the second elastomeric layer onto a top surface of the first elastomeric layer such that a control channel forms in the second recess between the first and second elastomeric layers; and positioning the first elastomeric layer on top of a planar substrate such that a flow channel forms in the first recess between the first elastomeric layer and the planar substrate. 1an elastomeric block; anda plurality of microfabricated channels formed in the elastomeric block, each channel containing a pressure or flow representing a signal, a change in the pressure or flow in a first channel resulting in a change in the pressure or fluid flow in a second channel consistent with a logic operation.. A fluidic logic device comprising: This nonprovisional patent application is a continuation on U.S. application Ser. No. 11/056,451 filed Feb. 10, 2005, which is a continuation of Ser. No. 09/826,583 filed Apr. 6, 2001, which is a continuation-in-part of U.S. application Ser. No. 09/724,784 filed Nov. 28, 2000, which is a continuation-in-part of U.S. application Ser. No. 09/605,520 filed Jun. 27, 2000, which claims the benefit of U.S. Appl. No. 60/141,503 filed Jun. 28, 1999; U.S. Appl. No. 60/147,199 filed Aug. 3, 1999; and U.S. Appl. No. 60/186,856 filed Mar. 3, 2000.Work described herein has been supported, in part, by National Institutes of Health grant HG-01642-02. The United States Government has certain rights in the invention.The present ...

System and Method for Microfluidic Cell Culture

Microfluidic devices and methods for perfusing a cell with perfusion fluid are provided herein, wherein the gravitational forces acting on the cell to keep the cell at or near a retainer or a retaining position exceed the hydrodynamic forces acting on the cell to move it toward an outlet. Also provided, are methods for assaying cell products within the microfluidic device.

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145.-. (canceled)46. A method for determining whether a secreted monoclonal antibody produced by a single antibody producing cell (APC) binds to an antigen , comprising:retaining a single APC within a chamber having a volume of from 100 μL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single APC within the chamber to produce a secreted monoclonal antibody;exposing the secreted monoclonal antibody to a first removeable capture substrate having an antigen on its surface, wherein the antigen is capable of binding the secreted monoclonal antibody, anddetermining whether the secreted monoclonal antibody binds the antigen.47. The method of claim 46 , wherein the single APC is a primary B cell or a memory B cell.48. The method of claim 46 , wherein the single APC is a primary plasma cell.49. The method of claim 46 , wherein the single APC is from a human claim 46 , a rabbit claim 46 , a rat claim 46 , a mouse claim 46 , a sheep claim 46 , an ape claim 46 , a monkey claim 46 , a goat claim 46 , a dog claim 46 , a cat claim 46 , a camel claim 46 , or a pig.50. The method of claim 46 , wherein the antigen is fluorescently labeled.51. The method of claim 50 , further comprising ...

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145.-. (canceled)46. A method of assaying a secreted monoclonal antibody produced by a single antibody producing cell (APC) and an antigen , comprising:retaining the single APC within a chamber having a volume of from 100 μL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single APC within the chamber to produce a secreted monoclonal antibody;exposing the secreted monoclonal antibody to a removeable capture substrate, wherein the removeable capture substrate is in fluid communication with the secreted monoclonal antibody and wherein the removeable capture substrate is capable of binding the secreted monoclonal antibody and nucleic acids of the single APC;incubating the secreted monoclonal antibody with the removeable capture substrate to produce a bound antibody;exposing a first fluid volume comprising the antigen in fluid communication with the bound antibody;determining whether the bound antibody binds the antigen; andlysing the single APC and capturing the nucleic acids of the single APC on the removeable capture substrate.47. The method of claim 46 , wherein the single APC is a primary B cell or a memory B cell.48. The method of claim 46 , wherein the single APC is a ...

Methods for Assaying Cellular Binding Interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule. 145.-. (canceled)46. A method of assaying a secreted monoclonal antibody produced by a single antibody producing cell (APC) , comprising:retaining the single APC within a chamber having a volume of from 100 pL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single APC within the chamber to produce a secreted monoclonal antibody;exposing the secreted monoclonal antibody to a first removeable capture substrate bound to an antigen;incubating the secreted monoclonal antibody with the removeable capture substrate; anddetermining whether the secreted monoclonal antibody binds the antigen.47. A method of assaying a secreted monoclonal antibody produced by a single antibody producing cell (APC) , comprising:retaining the single APC within a chamber having a volume of from 100 pL to 100 nL, a solid wall, and an aperture that defines an opening of the chamber;incubating the single APC within the chamber to produce a secreted monoclonal antibody;exposing the secreted monoclonal antibody to a first removeable capture substrate, wherein the first removeable capture substrate is in fluid communication with the secreted monoclonal antibody and is capable of binding the secreted ...

Microfluidic Devices and Methods for Use Thereof in Multicellular Assays of Secretion

Methods and devices are provided herein for identifying a cell population comprising an effector cell that exerts an extracellular effect. In one embodiment the method comprises retaining in a microreactor a cell population comprising one or more effector cells, wherein the contents of the microreactor further comprise a readout particle population comprising one or more readout particles, incubating the cell population and the readout particle population within the microreactor, assaying the cell population for the presence of the extracellular effect, wherein the readout particle population or subpopulation thereof provides a direct or indirect readout of the extracellular effect, and determining, based on the results of the assaying step, whether one or more effector cells within the cell population exerts the extracellular effect on the readout particle. If an extracellular effect is measured, the cell population is recovered for further analysis to determine the cell or cells responsible for the effect. 1218-. (canceled)219. A method of identifying an antibody secreting cell (ASC) that secretes a virus neutralizing antibody , comprising:retaining in a plurality of microreactors a plurality of cell populations, wherein individual cell populations of the plurality comprise one or more ASCs, wherein the contents of individual microreactors of the plurality of microreactors each comprise a readout particle population comprising one or more readout particles, and wherein the individual cell populations are retained in the individual microreactors;introducing a plurality of accessory particle populations into the individual microreactors, wherein individual accessory particle populations of the plurality comprise a plurality of virus particles and the individual accessory particle populations are retained in individual microreactors;incubating the individual cell populations, the readout particle populations and the accessory particle populations within the ...

System and Method for Microfluidic Cell Culture

Microfluidic devices and methods for perfusing a cell with perfusion fluid are provided herein, wherein the gravitational forces acting on the cell to keep the cell at or near a retainer or a retaining position exceed the hydrodynamic forces acting on the cell to move it toward an outlet. Also provided, are methods for assaying cell products within the microfluidic device.

Microfluidic Rotary Flow Reactor Matrix

A microfluidic device comprises a matrix of rotary flow reactors. The microfluidic matrix device offers a solution to the “world-to-chip” interface problem by accomplishing two important goals simultaneously: an economy of scale in reagent consumption is achieved, while simultaneously minimizing pipetting steps. N 2 independent assays can be performed with only 2N+1 pipetting steps, using a single aliquot of enzyme amortized over all reactors. The chip reduces labor relative to conventional fluid handling techniques by using an order of magnitude less pipetting steps, and reduces cost by consuming two to three orders of magnitude less reagents per reaction. A PCR format has immediate applications in medical diagnosis and gene testing. Beyond PCR, the microfluidic matrix chip provides a universal and flexible platform for biological and chemical assays requiring parsimonious use of precious reagents and highly automated processing.

Nucleic acid amplification using microfluidic devices

The present invention provides microfluidic devices and methods using the same in various types of thermal cycling reactions. Certaom devices include a rotary microfluidic channel and a plurality of temperature regions at different locations along the rotary microfluidic channel at which temperature is regulated. Solution can be repeatedly passed through the temperature regions such that the solution is exposed to different temperatures. Other microfluidic devices include an array of reaction chambers formed by intersecting vertical and horizontal flow channels, with the ability to regulate temperature at the reaction chambers. The microfluidic devices can be used to conduct a number of different analyses, including various primer extension reactions and nucleic acid amplification reactions.

Nucleic acid amplification using microfluidic devices

The present invention provides microfluidic devices and methods using the same in various types of thermal cycling reactions. Certain devices include a rotary microfluidic channel and a plurality of temperature regions at different locations along the rotary microfluidic channel at which temperature is regulated. Solution can be repeatedly passed through the temperature regions such that the solution is exposed to different temperatures. Other microfluidic devices include an array of reaction chambers formed by intersecting vertical and horizontal flow channels, with the ability to regulate temperature at the reaction chambers. The microfluidic devices can be used to conduct a number of different analyzes, including various primer extension reactions and nucleic acid amplification reactions.

Microfluidic rotary flow reactor matrix

A microfluidic device comprises a matrix of rotary flow reactors. The microfluidic matrix device offers a solution to the “world-to-chip” interface problem by accomplishing two important goals simultaneously: an economy of scale in reagent consumption is achieved, while simultaneously minimizing pipetting steps. N 2 independent assays can be performed with only 2N+1 pipetting steps, using a single aliquot of enzyme amortized over all reactors. The chip reduces labor relative to conventional fluid handling techniques by using an order of magnitude less pipetting steps, and reduces cost by consuming two to three orders of magnitude less reagents per reaction. A PCR format has immediate applications in medical diagnosis and gene testing. Beyond PCR, the microfluidic matrix chip provides a universal and flexible platform for biological and chemical assays requiring parsimonious use of precious reagents and highly automated processing.

Microfluidic rotary flow reactor matrix

A microfluidic device comprises a matrix of rotary flow reactors. The microfluidic matrix device offers a solution to the “world-to-chip” interface problem by accomplishing two important goals simultaneously: an economy of scale in reagent consumption is achieved, while simultaneously minimizing pipetting steps. N 2 independent assays can be performed with only 2N+1 pipetting steps, using a single aliquot of enzyme amortized over all reactors. The chip reduces labor relative to conventional fluid handling techniques by using an order of magnitude less pipetting steps, and reduces cost by consuming two to three orders of magnitude less reagents per reaction. A PCR format has immediate applications in medical diagnosis and gene testing. Beyond PCR, the microfluidic matrix chip provides a universal and flexible platform for biological and chemical assays requiring parsimonious use of precious reagents and highly automated processing.

Nucleic acid amplification utilizing microfluidic devices

Microfluidic rotary flow reactor matrix

A microfluidic device comprises a matrix of rotary flow reactors. The microfluidic matrix device offers a solution to the “world-to-chip” interface problem by accomplishing two important goals simultaneously: an economy of scale in reagent consumption is achieved, while simultaneously minimizing pipetting steps. N 2 independent assays can be performed with only 2N+1 pipetting steps, using a single aliquot of enzyme amortized over all reactors. The chip reduces labor relative to conventional fluid handling techniques by using an order of magnitude less pipetting steps, and reduces cost by consuming two to three orders of magnitude less reagents per reaction. A PCR format has immediate applications in medical diagnosis and gene testing. Beyond PCR, the microfluidic matrix chip provides a universal and flexible platform for biological and chemical assays requiring parsimonious use of precious reagents and highly automated processing.

Microfabricated elastomeric valve and pump systems

A method of fabricating an elastomeric structure, comprising: forming a first elastomeric layer on top of a first micromachined mold, the first micromachined mold having a first raised protrusion which forms a first recess extending along a bottom surface of the first elastomeric layer; forming a second elastomeric layer on top of a second micromachined mold, the second micromachined mold having a second raised protrusion which forms a second recess extending along a bottom surface of the second elastomeric layer; bonding the bottom surface of the second elastomeric layer onto a top surface of the first elastomeric layer such that a control channel forms in the second recess between the first and second elastomeric layers; and positioning the first elastomeric layer on top of a planar substrate such that a flow channel forms in the first recess between the first elastomeric layer and the planar substrate.

High throughput screening of crystallization of materials

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels.

Microfludic protein crystallography techniques

The present invention relates to microfluidic devices and methods facilitating the growth and analysis of crystallized materials such as proteins. In accordance with one embodiment, a crystal growth architecture is separated by a permeable membrane from an adjacent well having a much larger volume. The well may be configured to contain a fluid having an identity and concentration similar to the solvent and crystallizing agent employed in crystal growth, with diffusion across the membrane stabilizing that process. Alternatively, the well may be configured to contain a fluid having an identity calculated to affect the crystallization process. In accordance with the still other embodiment, the well may be configured to contain a material such as a cryo-protectant, which is useful in protecting the crystalline material once formed.

Microfluidic free interface diffusion techniques

A static fluid and a second fluid are placed into contact along a microfluidic free interface and allowed to mix by diffusion without convective flow across the interface. In accordance with one embodiment of the present invention, the fluids are static and initially positioned on either side of a closed valve structure in a microfluidic channel having a width that is tightly constrained in at least one dimension. The valve is then opened, and no-slip layers at the sides of the microfluidic channel suppress convective mixing between the two fluids along the resulting interface. Applications for microfluidic free interfaces in accordance with embodiments of the present invention include, but are not limited to, protein crystallization studies, protein solubility studies, determination of properties of fluidics systems, and a variety of biological assays such as diffusive immunoassays, substrate turnover assays, and competitive binding assays.

Microfluidic protein crystallography

The use of microfluidic structures enables high throughput screening of protein crystallization. In one embodiment, an integrated combinatoric mixing chip allows for precise metering of reagents to rapidly create a large number of potential crystallization conditions, with possible crystal formations observed on chip. In an alternative embodiment, the microfluidic structures may be utilized to explore phase space conditions of a particular protein crystallizing agent combination, thereby identifying promising conditions and allowing for subsequent focused attempts to obtain crystal growth.

Microfluidic protein crystallography techniques

The present invention relates to microfluidic devices and methods facilitating the growth and analysis of crystallized materials such as proteins. In accordance with one embodiment, a crystal growth architecture is separated by a permeable membrane from an adjacent well having a much larger volume. The well may be configured to contain a fluid having an identity and concentration similar to the solvent and crystallizing agent employed in crystal growth, with diffusion across the membrane stabilizing that process. Alternatively, the well may be configured to contain a fluid having an identity calculated to affect the crystallization process. In accordance with the still other embodiment, the well may be configured to contain a material such as a cryo-protectant, which is useful in protecting the crystalline material once formed.

Microfluidic chemostat

A chemostat is described that includes a growth chamber having a plurality of compartments. Each of the compartments may be fluidly isolated from the rest of the growth chamber by one or more actuatable valves. The chemostat may also include a nutrient supply-line to supply growth medium to the growth chamber, and an output port to remove fluids from the growth chamber. Also, a method of preventing biofilm formation in a growth chamber of a chemostat is described. The method may include the steps of adding a lysis agent to a isolated portion of the growth chamber, and reuniting the isolated portion with the rest of the growth chamber.

Microfluidic Sieve Valves

Sieve valves for use in micorfluidic device are provided. The valves are useful for impeding the flow of particles, such as chromatography beads or cells, in a microfluidic channel while allowing liquid solution to pass through the valve. The valves find particular use in making microfluidic chromatography modules.

Microfabricated elastomeric valve and pump systems

A method of fabricating an elastomeric structure, comprising: forming a first elastomeric layer on top of a first micromachined mold, the first micromachined mold having a first raised protrusion which forms a first recess extending along a bottom surface of the first elastomeric layer; forming a second elastomeric layer on top of a second micromachined mold, the second micromachined mold having a second raised protrusion which forms a second recess extending along a bottom surface of the second elastomeric layer; bonding the bottom surface of the second elastomeric layer onto a top surface of the first elastomeric layer such that a control channel forms in the second recess between the first and second elastomeric layers; and positioning the first elastomeric layer on top of a planar substrate such that a flow channel forms in the first recess between the first elastomeric layer and the planar substrate.

High throughput screening of crystallization of materials

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels.

High throughput screening of crystallization of materials

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels.

High throughput screening of crystallization materials

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels.

Microfluidic sieve valves

Sieve valves for use in microfluidic device are provided. The valves are useful for impeding the flow of particles, such as chromatography beads or cells, in a microfluidic channel while allowing liquid solution to pass through the valve. The valves find particular use in making microfluidic chromatography modules.

High throughput screening of crystallization of materials

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels.

Microfabricated elastomeric valve and pump systems

A method of fabricating an elastomeric structure, comprising: forming a first elastomeric layer on top of a first micromachined mold, the first micromachined mold having a first raised protrusion which forms a first recess extending along a bottom surface of the first elastomeric layer; forming a second elastomeric layer on top of a second micromachined mold, the second micromachined mold having a second raised protrusion which forms a second recess extending along a bottom surface of the second elastomeric layer; bonding the bottom surface of the second elastomeric layer onto a top surface of the first elastomeric layer such that a control channel forms in the second recess between the first and second elastomeric layers; and positioning the first elastomeric layer on top of a planar substrate such that a flow channel forms in the first recess between the first elastomeric layer and the planar substrate.

Methods for assaying cellular binding interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule.

Microfluidic chemostat

Microfluidic devices and methods for use thereof in multicellular assays of secretion

【課題】標的エピトープに結合する抗体を分泌する抗体分泌細胞（ＡＳＣ）を含む不均一細胞集団を同定するための方法の提供。【解決手段】１０〜５００の細胞を含む不均一細胞集団をマイクロリアクターに保持する工程であって、マイクロリアクターの中身は、標的エピトープを提示する１又は複数の読み出し粒子４１２、４１３、４１４を含む、読み出し粒子集団をさらに含む工程と、マイクロリアクター内で、不均一細胞集団と、読み出し粒子４１２、４１３、４１４とをインキュベートする工程であって、マイクロリアクターはその周囲の環境から実質的に隔離されている工程と、ＡＳＣから分泌された抗体４１９、４２３が不均一細胞集団中で標的エピトープに結合しているかをアッセイする工程と、及びアッセイする工程の結果に基づいて、不均一細胞集団が標的エピトープに結合する抗体４１９、４２３を分泌するＡＳＣを含むかどうかを決定する工程、を含む方法。【選択図】図２３

Microfluidic sieve valves

Sieve valves for use in micorfluidic device are provided. The valves are useful for impeding the flow of particles, such as chromatogrpahy beads or cells, in a microfluidic channel while allowing liquid solution to pass through the valve. The valves find particular use in making microfluidic chromatography modules.

Microfabricated elastomeric valve and pump systems

A method of fabricating an elastomeric structure, comprising: forming a first elastomeric layer on top of a first micromachined mold, the first micromachined mold having a first raised protrusion which forms a first recess extending along a bottom surface of the first elastomeric layer; forming a second elastomeric layer on top of a second micromachined mold, the second micromachined mold having a second raised protrusion which forms a second recess extending along a bottom surface of the second elastomeric layer; bonding the bottom surface of the second elastomeric layer onto a top surface of the first elastomeric layer such that a control channel forms in the second recess between the first and second elastomeric layers; and positioning the first elastomeric layer on top of a planar substrate such that a flow channel forms in the first recess between the first elastomeric layer and the planar substrate.

Microfluidic devices and methods for use thereof in multicellular assays of secretion

Methods and devices are provided herein for identifying a cell population comprising an effector cell that exerts an extracellular effect. In one embodiment the method comprises retaining in a microreactor a cell population comprising one or more effector cells, wherein the contents of the microreactor further comprise a readout particle population comprising one or more readout particles, incubating the cell population and the readout particle population within the microreactor, assaying the cell population for the presence of the extracellular effect, wherein the readout particle population or subpopulation thereof provides a direct or indirect readout of the extracellular effect, and determining, based on the results of the assaying step, whether one or more effector cells within the cell population exerts the extracellular effect on the readout particle. If an extracellular effect is measured, the cell population is recovered for further analysis to determine the cell or cells responsible for the effect.

Microfluidic protein crystallography

The use of microfluidic structures enables high throughput screening of protein crystallization. In one embodiment, an integrated combinatoric mixing chip allows for precise metering of reagents to rapidly create a large number of potential crystallization conditions, with possible crystal formations observed on chip. In an alternative embodiment, the microfluidic structures may be utilized to explore phase space conditions of a particular protein crystallizing agent combination, thereby identifying promising conditions and allowing for subsequent focused attempts to obtain crystal growth.

Microfluidic protein crystallography

Microfluidic protein crystallography

The use of microfluidic structures enables high throughput screening of protein crystallization. In one embodiment, an integrated combinatoric mixing chip allows for precise metering of reagents to rapidly create a large number of potential crystallization conditions, with possible crystal formations observed on chip. In an alternative embodiment, the microfluidic structures may be utilized to explore phase space conditions of a particular protein crystallizing agent combination, thereby identifying promising conditions and allowing for subsequent focused attempts to obtain crystal growth.

Microfluidic devices and methods for use thereof in multicellular assays of secretion

Microfluidic protein crystallography

The use of microfluidic structures enables high throughput screening of protein crystallization. In one embodiment, an integrated combinatoric mixing chip allows for precise metering of reagents to rapidly create a large number of potential crystallization conditions, with possible crystal formations observed on chip. In an alternative embodiment, the microfluidic structures may be utilized to explore phase space conditions of a particular protein crystallizing agent combination, thereby identifying promising conditions and allowing for subsequent focused attempts to obtain crystal growth.

Methods for assaying cellular binding interactions

There are provided methods, and devices for assaying for a binding interaction between a protein, such as a monoclonal antibody, produced by a cell, and a biomolecule. The method may include retaining the cell within a chamber having an aperture; exposing the protein produced by the cell to a capture substrate, wherein the capture substrate is in fluid communication with the protein produced by the cell and wherein the capture substrate is operable to bind the protein produced by the cell; flowing a fluid volume comprising the biomolecule through the chamber via said aperture, wherein the fluid volume is in fluid communication with the capture substrate; and determining a binding interaction between the protein produced by the cell and the biomolecule.

Microfluidic devices and methods for use thereof in multicellular assays of secretion

Methods and devices are provided herein for identifying a cell population comprising an effector cell that exerts an extracellular effect. In one embodiment the method comprises retaining in a microreactor a cell population comprising one or more effector cells, wherein the contents of the microreactor further comprise a readout particle population comprising one or more readout particles, incubating the cell population and the readout particle population within the microreactor, assaying the cell population for the presence of the extracellular effect, wherein the readout particle population or subpopulation thereof provides a direct or indirect readout of the extracellular effect, and determining, based on the results of the assaying step, whether one or more effector cells within the cell population exerts the extracellular effect on the readout particle. If an extracellular effect is measured, the cell population is recovered for further analysis to determine the cell or cells responsible for the effect.

Microfluidic devices and methods for use thereof in multicellular assays of secretion

Methods and devices are provided herein for identifying a cell population comprising an effector cell that exerts an extracellular effect. In one embodiment the method comprises retaining in a microreactor a cell population comprising one or more effector cells, wherein the contents of the microreactor further comprise a readout particle population comprising one or more readout particles, incubating the cell population and the readout particle population within the microreactor, assaying the cell population for the presence of the extracellular effect, wherein the readout particle population or subpopulation thereof provides a direct or indirect readout of the extracellular effect, and determining, based on the results of the assaying step, whether one or more effector cells within the cell population exerts the extracellular effect on the readout particle. If an extracellular effect is measured, the cell population is recovered for further analysis to determine the cell or cells responsible for the effect.

分泌物の多細胞アッセイにおけるマイクロ流体デバイスおよびその使用方法

【課題】ウイルスを中和する抗体を産生する抗体分泌細胞を同定する方法を提供する。 【解決手段】方法は、複数のマイクロリアクターに、個々の細胞集団が１つ又は複数の抗体分泌細胞を含む複数の細胞集団を保持する工程と、１つ又は複数の読み出し粒子を感染するように動作可能な複数のウイルス粒子を含む、複数のアクセサリー粒子集団を該複数のマイクロリアクターに導入する工程と、該個々の細胞集団、該読み出し粒子、および該アクセサリー粒子集団を、複数の抗体を産生するのに十分な時間、該個々のマイクロリアクター内でインキュベートする工程と、該個々のマイクロリアクターをアッセイし、該複数の抗体が該ウイルスを中和する抗体を含むかどうかを決定する工程と、及び該ウイルスを中和する抗体を産生する抗体分泌細胞を同定する工程と、を含む。 【選択図】図２３

Microfluidic devices and methods for use thereof in multicellular assays of secretion

Methods and devices are provided herein for identifying a cell population comprising an effector cell that exerts an extracellular effect. In one embodiment the method comprises retaining in a microreactor a cell population comprising one or more effector cells, wherein the contents of the microreactor further comprise a readout particle population comprising one or more readout particles, incubating the cell population and the readout particle population within the microreactor, assaying the cell population for the presence of the extracellular effect, wherein the readout particle population or subpopulation thereof provides a direct or indirect readout of the extracellular effect, and determining, based on the results of the assaying step, whether one or more effector cells within the cell population exerts the extracellular effect on the readout particle. If an extracellular effect is measured, the cell population is recovered for further analysis to determine the cell or cells responsible for the effect.

Dough brake