Microfabricated crossflow devices and methods

• US 8,252,539 B2
• Filed: 10/08/2007
• Issued: 08/28/2012
• Est. Priority Date: 09/15/2000
• Status: Active Grant

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Quake, Stephen R., Thorsen, Todd
• Primary Examiner(s): Beisner, William H
• Application Number:
  US11/868,942
• Publication Number:
  US 20090035838A1
• Time in Patent Office: 1,786 Days
• Field of Search: 435/288.5
• US Class Current: 435/6.12
• 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 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/0418 electro-osmotic flow [EOF]
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0454 radiation pressure, optical tweezers
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0484 Cantilevers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0638 membrane valves, flap valves
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: C12M 1/34 : Measuring or testing with condition measuring or sensing means, e.g. colony counters
• CPC Class Codes: C12Q 1/68 : involving nucleic acids
• CPC Class Codes: G01N 15/01 : specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/1404 : Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1434 : Optical arrangements
• CPC Class Codes: G01N 15/1459 : the analysis being performed on a sample stream
• 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 15/1492 : within droplets
• CPC Class Codes: G01N 2015/019 : Biological contaminants; Fouling
• CPC Class Codes: G01N 2015/1006 : for cytology
• CPC Class Codes: G01N 2015/1028 : Sorting particles
• CPC Class Codes: G01N 2015/1411 : Features of sheath fluids
• CPC Class Codes: G01N 2015/1481 : Optical analysis of particles within droplets sorting particles within droplets G01N15/1492
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: Y10T 436/118339 : with formation of a segmented stream
• CPC Class Codes: Y10T 436/143333 : Saccharide [e.g., DNA, etc.]

Abstract

A microfluidic device for analyzing and/or sorting biological materials (e.g., molecules such as polynucleotides and polypeptides, including proteins and enzymes; viruses and cells) and methods for its use are provided. The device and methods of the invention are useful for sorting particles, e.g. virions. The invention is also useful for high throughput screening, e.g. combinatorial screening. The microfluidic device comprises a main channel and an inlet region in communication with the main channel at a droplet extrusion region. Droplets of solution containing the biological material are deposited into the main channel through the droplet extrusion region. A fluid different from and incompatible with the solution containing the biological material flows through the main channel so that the droplets containing the biological material do not diffuse or mix. Biological material within the droplets can be analyzed and/or sorted by detecting a predetermined characteristic of the biological sample in each droplet and sorting the droplet accordingly.

Claims

1. A method of making droplets in a microfluidic device, comprising flowing an extrusion fluid through a first input channel at a first pressure while flowing a sample fluid through a second input channel at a second pressure,

wherein the sample fluid is immiscible with the extrusion fluid, wherein the first input channel and the second input channel are in fluidic communication at a junction, wherein the junction is constructed and arranged so that sample fluid droplets are sheared into an output channel,

wherein the first and second input channels and the output channel have diameters between about 2 and 100 microns or cross-sectional dimensions in the range of 1 to 100 microns.

2. The method of claim 1, wherein the first input channel, the second input channel, and the output channel have diameters that range between 2 and 100 microns.

3. The method of claim 1, wherein the first pressure is higher than the second pressure.

4. The method of claim 1, wherein the first pressure is approximately equal to the second pressure.

5. The method of claim 1, wherein the first pressure is less than the second pressure.

6. The method of claim 1, wherein the relative pressure between the first pressure and the second pressure is selected so as to produce monodisperse sample fluid droplets having regular periodicity.

7. The method of claim 1, wherein a length dimension of the sample fluid droplets is longer than a width dimension of the sample fluid droplets.

8. The method of claim 1, wherein the sample fluid droplets introduced into the extrusion fluid are monodisperse.

9. The method of claim 1, wherein the sample fluid droplets are packed in ordered layers in the output channel.

10. The method of claim 1, wherein the sample fluid droplets contain a biological material.

11. The method of claim 10, wherein the sample fluid droplets contain a polynucleotide and/or an enzyme.

12. The method of claim 11, wherein the sample fluid droplets contain a reporter molecule.

13. The method of claim 12, wherein the reporter molecule is a fluorescent agent.

14. The method of claim 11, further comprising detecting signal produced by a chemical reaction of a substrate catalyzed by an enzyme in the sample fluid droplets.

15. The method of claim 11, further comprising detecting signal produced by a polymerase chain reaction in the sample fluid droplets.

16. The method of claim 10, wherein the sample fluid droplets contain, on average, no more than one particle of the biological material.

17. The method of claim 1, wherein the sample fluid droplets contain a sample of molecules for small scale chemical reactions.