Microfluidic particle-analysis systems

• US 8,658,418 B2
• Filed: 07/13/2009
• Issued: 02/25/2014
• Est. Priority Date: 04/01/2002
• 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:
  US12/501,982
• Publication Number:
  US 20100120077A1
• Time in Patent Office: 1,688 Days
• Field of Search: 435/287.2, 435/288.3, 435/288.4, 435/288.5, 435/305.1, 435/305.2
• US Class Current: 435/288.5
• 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 analysis. 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 analysis, 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 device for exposing cells to a plurality of compounds comprising:

(a) a cell inlet;

(b) a plurality of cell chambers in fluidic communication with said cell inlet and at least one of the other cell chambers;

(c) a plurality of chamber isolation valves, said isolation valves, when actuated, fluidically isolating said chambers from each other;

(d) a plurality of compound inlets and compound outlets, wherein each chamber is in fluidic communication with a compound inlet and a compound outlet, wherein each chamber includes a channel sized for retaining a single cell within said chamber;

(e) a shared waste receptacle, said shared waste receptacle being configured to receive waste from said compound outlets;

(f) a shared reagent reservoir, said shared reagent reservoir being configured to provide reagent to said chambers;

(g) a plurality of compound outlet valves, said compound outlet valves configured to fluidically isolate said chambers from said shared waste receptacle and said shared reagent reservoir when closed;

(h) a plurality of analysis sites, wherein each cell chamber has a corresponding analysis site, wherein each chamber is in fluidic communication with a respective analysis site via its channel;

wherein the plurality of cell chambers includes a first cell chamber and a second cell chamber downstream from the first cell chamber,

wherein the first cell chamber is configured to hold a first cell using a retaining force fluidically applied via its channel and thereby allow a second cell following the first cell to be diverted downstream along a shared cell input channel to the second cell chamber.

2. The device of claim 1 wherein at least one of said plurality of compounds is a cell toxic compound.

3. The device of claim 1 wherein said common reagent reservoir contains a detection agent responsive to a toxic response of said cells to one of said compounds.

4. The device of claim 1, wherein the plurality of cell chambers is arranged such that an entrance of each cell chamber following the first cell chamber is sequentially arranged downstream to the entrance of another cell chamber.

5. The device of claim 1, wherein the first cell chamber is physically sized to hold only a single cell according a size range encompassing each of the first and second cells, and thereby configured to only hold the first cell, which blocks the second cell from coupling to the first cell chamber.

6. The device of claim 1, wherein the fluidic retaining force fluidically applied to the channel prevents backflow of the first cell into the shared cell input channel while the second cell is flowed within the input channel.

7. The device of claim 1, wherein each chamber is in permanent fluidic communication with its respective analysis site via its channel.

8. The device of claim 1, wherein each channel comprises a switchable filter.

9. A method for determining whether one of a plurality of compounds causes a toxic response in one of a plurality of cells comprising the steps of:

providing the device of claim 1;

introducing said cells into said device of claim 1,

isolating said cells within said chambers of said device of claim 1,

exposing said cells to said plurality of said compounds while said cells are retained in said chambers,

determining whether any of said cells had a toxic response resulting from exposure to one of said plurality of compounds.