Microfluidic particle-analysis systems

• US 7,312,085 B2
• Filed: 04/01/2003
• Issued: 12/25/2007
• Est. Priority Date: 04/01/2002
• Status: Active Grant

• Current Assignee: Standard Biotools Incorporated
• Original Assignee: Fluidigm Corporation
• Inventors: Chou, Hou-Pu, Daridon, Antoine, Farrell, Kevin, Fowler, Brian, Liau, Yish-Hann, Manger, Ian D., Nassef, Hany Ramez, Throndset, William
• Primary Examiner(s): SINES, BRIAN J
• Application Number:
  US10/405,953
• Publication Number:
  US 20040072278A1
• Time in Patent Office: 1,729 Days
• Field of Search: 422/50, 422/68.1, 422/81, 422/82, 422/100, 422/101, 422/102, 422/103, 422/104, 436/43, 436/63, 436/180, 435/4, 435/404, 435/405, 435/325, 435/382, 435/383, 435/384
• US Class Current: 436/43
• 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 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 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0655 pinch valves
• 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: G01N 15/1456 without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
• CPC Class Codes: G01N 15/1468 : with spatial resolution of the texture or inner structure of the particle
• 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 2015/0288 : Sorting the particles
• CPC Class Codes: G01N 2015/1027 : Determining speed or velocity of a particle
• CPC Class Codes: G01N 2015/1493 : Particle size
• CPC Class Codes: G01N 35/00 : Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

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 method for treating a particle comprising the steps of:

(i) providing a microfluidic device comprising;

an input mechanism for introducing a fluid sample containing a particle;

a microfluidic passage in fluid communication with said input mechanism;

a positioning mechanism using a reduced-velocity flow stream in fluid communication with said microfluidic passage, said positioning mechanism for positioning said particle in said microfluidic passage while contained in said fluid sample;

a flow-assisted retention mechanism for retaining said particle upon being positioned by said positioning means;

a treatment mechanism in communication with said retention mechanism for selectively treating said particle 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 particle,

(ii) introducing said sample fluid containing said particle into said input mechanism;

(iii) positioning said particle with said positioning mechanism using a reduced-velocity flow stream so that said particle is retainable by said flow-assisted etention mechanism;

(iv) retaining said particle with said flow-assisted retention mechanism;

(v) exposing said particle to said treatment by said treatment mechanism;

(vi) measuring said treatment response caused directly or indirectly by said particle upon exposure to said treatment.

2. The method of claim 1 wherein said microfluidic device further comprises a release mechanism for releasing said particle from said flow-assisted retention mechanism, and said method further comprises the step of releasing said particle from said flow-assisted retention mechanism.

3. The method of claim 2, wherein said microfluidic device further comprises an output mechanism for outputting said particle from said microfluidic device, and said method further comprises the step of outputting said particle from said microfluidic device by said output mechanism.

4. The method of claim 2, wherein said microfluidic device further comprises a cell culture mechanism for culturing said particle, and the method further comprises the step of culturing said particle in said cell culture mechanism.

5. The method of claim 1, wherein said microfluidic device further comprises a control mechanism for determining aspects of the flow rate or path of the sample fluid or other fluid, and the method further comprises the step of determining the flow rate or path of the sample fluid or other fluid by said control mechanism.

6. The method of claim 5, wherein said control mechanism is a valve in communication with said microfluidic passage, and the method further comprises valving said sample fluid or other fluid with said valve.

7. The microfluidic device of claim 6, wherein said microfluidic device is formed from a multi-layer elastomeric block and, wherein said valve is formed from an elastomeric membrane within said elastomeric block, and wherein said valving occurs by deflecting said elastomeric membrane into said microfluidic passage.

8. The method of claim 6, wherein said control mechanism is a pump in communication with said microfluidic passage, and wherein said determining the flow rate or path of said sample fluid occurs by actuation of said pump.

9. The method of claim 8, wherein said microfluidic device is formed from a multi-layer elastomeric block and, wherein said pump is formed from an elastomeric membrane within said elastomeric block, and wherein said pump is actuated by deflecting a series of elastomeric membranes into said microfluidic passage in a selected sequence.

10. The method of claim 1, wherein said microfluidic device comprises a multi-layered elastomeric block having a control layer having an elastomeric membrane deflectable into said microfluidic passage in a fluidic layer to selectively determine the flow rate or path of a fluid in said microfluidic passage.

11. The method of claim 1, wherein said microfluidic passage further comprises an adjacent passage joining said microfluidic passage at a junction or branch, said adjacent passage being selected from the group consisting of inlet passage, outlet passage, particle passage, reagent passage, and waste passage, and said method further comprises the step of selectively determining the path of said particle to said adjacent passage.

12. The method of claim 11, wherein said adjacent passage is a dead-end passage, and wherein said selectively determining includes introducing said sample fluid into said dead-end passage wherein said sample fluid displaces gas, if present, in said dead-end passage to fill said dead-end passage with said sample fluid.

13. The method of claim 11 further comprising said adjacent passage manipulating said particle.

14. The method of claim 13, wherein said particle manipulating includes retaining said particle in addition to either positioning, sorting, treating, detecting, propagating, storing, mixing, or releasing said particle.

15. The method of claim 1, wherein said particle is selected from the group consisting of cells, eukaryotic cells, prokaryotic cells, plant cells, animal cells, hybridoma cells, bacterial cells, yeast cells, viruses, organelles, beads, and vesicles, and wherein said treating step treats said particle.

16. The method of claim 15, wherein said particle is a plurality or an aggregate of particles, and said method further comprises a sorting step to sort out and separate or isolate a desired particle from said plurality of particles, and said treating step treats said separated or isolated particle.

17. The method of claim 16, wherein said plurality of particles is a complex mixture containing different particles, and said sorting step sorts out at least one type of particle from other different particles in said complex mixture.

18. The method of claim 17, wherein said complex mixture containing different particles is whole blood or serum or bodily fluid, and said sorting step selects for at least one type of cell from the whole blood or serum.

19. The method of claim 1, wherein said particle is an egg or embryo, and said treatment is a step towards in-vitro fertilizing or manipulating said egg or embryo, respectively.

20. The method of claim 1, wherein the input mechanism is a receptacle or well in fluid communication with said microfluidic passage, and said method further comprises the step of introducing said fluid sample into said receptacle.