Microfluidic particle-analysis systems

• US 7,452,726 B2
• Filed: 12/11/2003
• Issued: 11/18/2008
• Est. Priority Date: 04/01/2002
• Status: Expired due to Fees

• Current Assignee: Fluidigm Corporation
• Original Assignee: Fluidigm Corporation
• Inventors: Manger, Ian D., Daridon, Antoine, Chou, Hou-Pu, Farrell, Kevin, Nassef, Hany Ramez, Fowler, Brian, Liau, Yish-Hann, Throndset, William
• Primary Examiner(s): WARDEN, JILL ALICE
• Application Number:
  US10/734,963
• Publication Number:
  US 20040224380A1
• Time in Patent Office: 1,804 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, 435/325, 435/382, 435/383, 435/384, 435/404, 435/405
• US Class Current: 436/63
• 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 perfusing cells with a reagent comprising the steps of:

(a) providing a microfluidic device having

(i) a cell growth chamber,

a cell inlet in communication with said chamber, said cell inlet having an inlet valve in operable communication therewith to control fluid flow through said cell inlet into said chamber, wherein said cells can pass through said cell inlet into said chamber when said inlet valve is open, but cannot pass through said cell inlet when said inlet valve is closed; and

,

(ii) a reagent inlet for inputting said reagent into said chamber, said reagent inlet having a reagent valve in operable communication with said reagent inlet for controlling fluid flow through said reagent inlet into said chamber, said cell inlet and/or said chamber having a retention mechanism for retaining said cells in said chamber while permitting flow of said reagent into said chamber when said reagent valve is open;

wherein when said cells are loaded into said chamber, and said inlet valve is closed, said cells are retained in said chamber by the retention mechanism while said reagent valve is open and closed;

(b) opening said cell inlet valve and introducing said cells into said chamber;

(c) closing said cell inlet valve;

(d) opening said reagent valve to introduce said reagent into said chamber; and

,

(e) introducing said reagent into said chamber while retaining said cells inside of said chamber thereby perfusing said cells with said reagent.

2. The method recited in claim 1 wherein the microfluidic device further comprises a release mechanism for releasing the cells from the retention mechanism, the method further comprising releasing the cells from the retention mechanism.

3. The method recited in claim 2 wherein the microfluidic device further comprises an output mechanism for outputting the cells from the microfluidic device, the method further comprising outputting the cells from the microfluidic device with the output mechanism.

4. The method recited in claim 2 wherein the microfluidic device further comprises a cell culture mechanism for culturing the cells, the method further comprising culturing the cells in the cell culture mechanism.

5. The method recited in claim 1 wherein:

the microfluidic device comprises a multi-layered elastomeric block having a control layer; and

the inlet valve comprises an elastomeric membrane comprised by the control layer and deflectable into an elastomeric passage in a fluid layer to selectively determine a flow rate or flow path of a fluid in the elastomeric passage.

6. The method recited in claim 1 wherein the microfluidic device further comprises a treatment mechanism in communication with the retention mechanism, the method further comprising selectively treating at least one of the cells with the treatment mechanism to produce a treatment response while the at least one of the cells is retained within the retention mechanism.

7. The method recited in claim 6 wherein the cells are selected from the group consisting of eukaryotic cells, prokaryotic cells, plants cells, animal cells, hybridoma cells, bacterial cells, and yeast cells.

8. The method recited in claim 7 further comprising isolating the at least one of the cells from a remainder of the cells.

9. The method recited in claim 7 wherein:

the at least one of the cells comprises an egg or an embryo; and

treating the at least one of the cells comprises a step towards in vitro fertilizing of the egg or manipulating the embryo.

10. The method recited in claim 1 wherein the cell inlet comprises a receptacle or well in fluid communication with the chamber, the method further comprising introducing fluid containing the cells into the receptacle or well.