Microfluidic particle-analysis systems

• US 9,926,521 B2
• Filed: 02/24/2014
• Issued: 03/27/2018
• Est. Priority Date: 06/27/2000
• Status: Expired due to Term

• Current Assignee: Standard Biotools Incorporated
• Original Assignee: Fluidigm Corporation
• Inventors: Daridon, Antoine
• Primary Examiner(s): BOWERS, NATHAN ANDREW
• Application Number:
  US14/188,660
• Publication Number:
  US 20140378352A1
• Time in Patent Office: 1,492 Days
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/06 Auxiliary integrated devices, integrated components
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/088 Channel loops
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/0893 having a very large number of wells, microfabricated wells
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0622 : distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502746 : characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502753 : characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12M 1/34 : Measuring or testing with condition measuring or sensing means, e.g. colony counters
• CPC Class Codes: C12M 21/06 : for in vitro fertilization
• CPC Class Codes: C12M 23/16 : Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: G01N 15/1484 : microstructural devices
• CPC Class Codes: G01N 15/149 : specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/016 : White blood cells
• CPC Class Codes: G01N 2015/0288 : Sorting the particles
• CPC Class Codes: G01N 2015/1493 : Particle size
• CPC Class Codes: G01N 33/4833 : of solid biological material, e.g. tissue samples, cell cultures tissue in vivo A61B5/00; cell suspensions G01N33/48735
• CPC Class Codes: G02B 21/32 : Micromanipulators structurally combined with microscopes

Abstract

The invention provides systems, including apparatus, methods, and kits, for the microfluidic manipulation and/or detection of particles, such as cells and/or beads. The invention provides systems, including apparatus, methods, and kits, for the microfluidic manipulation and/or analysis of particles, such as cells, viruses, organelles, beads, and/or vesicles. The invention also provides microfluidic mechanisms for carrying out these manipulations and analyses. These mechanisms may enable controlled input, movement/positioning, retention/localization, treatment, measurement, release, and/or output of particles. Furthermore, these mechanisms may be combined in any suitable order and/or employed for any suitable number of times within a system. Accordingly, these combinations may allow particles to be sorted, cultured, mixed, treated, and/or assayed, among others, as single particles, mixed groups of particles, arrays of particles, heterogeneous particle sets, and/or homogeneous particle sets, among others, in series and/or in parallel. In addition, these combinations may enable microfluidic systems to be reused. Furthermore, these combinations may allow the response of particles to treatment to be measured on a shorter time scale than was previously possible. Therefore, systems of the invention may allow a broad range of cell and particle assays, such as drug screens, cell characterizations, research studies, and/or clinical analyses, among others, to be scaled down to microfluidic size. Such scaled-down assays may use less sample and reagent, may be less labor intensive, and/or may be more informative than comparable macrofluidic assays.

Claims

1. A microfluidic system for cell manipulation comprising:

(a) an input mechanism for introducing a fluid sample containing a plurality of eukaryotic cells;

(b) a microfluidic passage in fluid communication with said input mechanism;

(c) a flow-based positioning mechanism in fluid communication with said microfluidic passage, said positioning mechanism for positioning at least one cell of the plurality of cells in a retention mechanism while contained in said fluid sample;

wherein the retention mechanism is for retaining one individual cell upon being positioned by said positioning mechanism,

wherein said retention mechanism is configured to retain a single cell and comprises a physical barrier that restricts longitudinal cell movement along the direction of fluid flow and restricts orthogonal cell movement, and

wherein the flow-based positioning mechanism comprises a dividing flow site formed by a junction of the microfluidic passage, a first outlet channel, and a second outlet channel, such that when the fluid sample containing the plurality of eukaryotic cells is introduced, said one individual cell is delivered to and retained by the retention mechanism, at least some unretained cells flow through the first outlet channel, and at least some unretained cells flow through the second outlet channel; and

,

(d) a release mechanism for releasing said cell from said retention mechanism.

2. The microfluidic system of claim 1 wherein the flow-based positioning mechanism comprises a dividing flow site formed by a junction of the microfluidic passage, a first outlet channel, and a second outlet channel, such that when the fluid sample containing the plurality of eukaryotic cells is introduced, said one individual cell is delivered to and retained by the retention mechanism, at least some unretained cells flow through the first outlet channel, and at least some unretained cells flow through the second outlet channel, wherein said individual cell, when retained by said retention mechanism, blocks fluid flow through the retention mechanism so that the retention mechanism preferentially retains only one cell.

3. The microfluidic system of claim 1 wherein said release mechanism is for terminating the flow-based positioning mechanism and releasing said cell from said retention mechanism.

4. The microfluidic system of claim 1 wherein said cell, when retained by said retention mechanism, blocks fluid flow through the retention mechanism so that the retention mechanism preferentially retains only one cell.

5. The microfluidic system of claim 4 wherein said release mechanism is for rendering ineffective the retention mechanism by lysing said cell to release intracellular components.

6. The microfluidic system of claim 1, wherein said microfluidic device comprises a layer including a material selected from the group consisting of elastomers, polydimethylsiloxane, plastic, polystyrene, polypropylene, polycarbonate, glass, ceramic, silicon, sol-gels, metal, metalloids, metal oxides, biological polymers, mixtures thereof, particles, proteins, gelatins, polylysine, serum albumin, collagen, nucleic acids, and microorganisms.

7. The microfluidic system of claim 1 further comprising a treatment mechanism in communication with said retention mechanism for selectively treating said cell to produce a treatment response while being retained within said retention mechanism;

and, a measurement mechanism for measuring said treatment response, if any, of said cell while said cell is stationary.

8. The microfluidic system of claim 7, wherein said treatment mechanism exposes said particle to a reagent or physical condition.

9. The microfluidic system of claim 7 wherein the fluid sample contains a eukaryotic cell that is labeled with a dye, the treatment mechanism is a light source for illuminating the labeled cell to produce a detectable signal while said labeled cell is stationary and being retained by said retention mechanism;

and the measurement mechanism is for measuring said detectable signal, if any, emitted by said labeled cell in response to the illumination while said labeled cell is stationary.

10. The microfluidic system of claim 9, wherein said dye is a fluorophore and the measurement mechanism is a detector associated with said microfluidic device that detects fluorescence.

11. A microfluidic system for cell manipulation comprising:

(a) an input mechanism for introducing a fluid sample containing a plurality of eukaryotic cells;

(b) a microfluidic passage in fluid communication with said input mechanism;

(c) a flow-based positioning mechanism in fluid communication with said microfluidic passage, said positioning mechanism for positioning at least one cell of the plurality of cells in a retention mechanism while contained in said fluid sample;

wherein the retention mechanism is for retaining one individual cell upon being positioned by said positioning mechanism,

wherein said retention mechanism is configured to retain a single cell and comprises a physical barrier that restricts longitudinal cell movement along the direction of fluid flow and restricts orthogonal cell movement, and

wherein the flow-based positioning mechanism comprises a dividing flow site formed by a junction of the microfluidic passage, a first outlet channel, and a second outlet channel, such that when the fluid sample containing the plurality of eukaryotic cells is introduced, said one individual cell is delivered to and retained by the retention mechanism, at least some unretained cells flow through the first outlet channel, and at least some unretained cells flow through the second outlet channel; and

,

(d) a release mechanism for releasing said cell from said retention mechanism,

wherein said release mechanism is for rendering ineffective the retention mechanism by lysing said single cell to release intracellular components.

12. The microfluidic system of claim 11 wherein said single cell, when retained by said retention mechanism, blocks fluid flow through the retention mechanism so that the retention mechanism preferentially retains only one cell.

13. The microfluidic system of claim 11, wherein said microfluidic device comprises a layer including a material selected from the group consisting of elastomers, polydimethylsiloxane, plastic, polystyrene, polypropylene, polycarbonate, glass, ceramic, silicon, sol-gels, metal, metalloids, metal oxides, biological polymers, mixtures thereof, particles, proteins, gelatins, polylysine, serum albumin, collagen, nucleic acids, and microorganisms.

14. The microfluidic system of claim 11, wherein said microfluidic passage is less than about 500 micrometers wide.

15. The microfluidic system of claim 11 further comprising a treatment mechanism in communication with said retention mechanism for selectively treating said single cell to produce a treatment response while being retained within said retention mechanism;

and, a measurement mechanism for measuring said treatment response, if any, of said cell while said cell is stationary.

16. The microfluidic system of claim 15 wherein the fluid sample contains a eukaryotic cell that is labeled with a dye, the treatment mechanism is a light source for illuminating the labeled cell to produce a detectable signal while said labeled cell is stationary and being retained by said retention mechanism;

and the measurement mechanism is for measuring said detectable signal, if any, emitted by said labeled cell in response to the illumination while said labeled cell is stationary.

17. The microfluidic system of claim 16, wherein said dye is a fluorophore and the measurement mechanism is a detector associated with said microfluidic device that detects fluorescence.

18. The microfluidic system of claim 15, wherein said treatment mechanism exposes said single cell to a reagent or physical condition.