Microfabricated crossflow devices and methods

• US 8,592,215 B2
• Filed: 09/29/2011
• Issued: 11/26/2013
• Est. Priority Date: 09/15/2000
• Status: Expired due to Term

• 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:
  US13/249,212
• Publication Number:
  US 20120231444A1
• Time in Patent Office: 789 Days
• Field of Search: None
• US Class Current: 436/53
• 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

This invention provides microfabricated devices and methods for detecting, analyzing and sorting biological materials and particles. Droplets containing the particles are provided in an extrusion fluid, passed through a detection region, and then directed into a branch channel according to predetermined characteristics. For example, cells or viral particles contained in droplets of aqueous solvent are flowed past a detector in the nonpolar extrusion fluid decane, and routed into a selected branch channel for subsequent analysis or use.

Claims

1. A method for sorting particles in a microfluidic device, the method comprising:

(a) flowing a sample fluid containing particles through a first channel while flowing an extrusion fluid immiscible with the sample fluid through a second channel towards a junction of the two channels that is configured so that droplets of the sample fluid are introduced into the extrusion fluid;

(b) flowing the droplets formed in step (a) through a detection region that is downstream from the junction;

(c) interrogating particle(s) in each droplet while the droplet is flowing through the detection region; and

(d) directing each droplet into a branched channel that has been selected from two or more branched channels depending on results of the interrogating in step (c).

2. The method of claim 1 wherein the-particles are viral particles.

3. The method of claim 2 wherein each of the droplets contains, on average, no more than one viral particle.

4. The method of claim 1 wherein:

the sample fluid is an aqueous solution; and

the extrusion fluid is a non-polar solvent.

5. The method of claim 4 wherein the aqueous solution comprises ultra pure water, TE buffer, phosphate buffered saline, or acetate buffer.

6. The method of claim 4 wherein the extrusion fluid is decane.

7. The method of claim 4 wherein the extrusion fluid is selected from tetradecane and hexadecane.

8. The method of claim 1 wherein each of the droplets contains, on average, no more than one of said particles.

9. The method of claim 1, wherein the refractive index of the extrusion fluid is different from the refractive index of the sample fluid, and wherein step (d) comprises the application of radiation pressure.

10. The method of claim 9, wherein the application of radiation pressure comprises moving droplets with an optical beam.

11. A method for sorting droplets in a microfluidic device, the device comprising:

(i) a main channel;

(ii) an inlet channel that forms a junction with the main channel;

(iii) a detection region in the main channel that is downstream from the junction and associated with a detector;

(iv) at least two branched channels downstream from the detection region;

(v) a flow control system responding to the detector and configured to direct individual droplets into one of said branched channels;

the method comprising;

(a) flowing a sample fluid through the inlet channel to the junction while flowing extrusion fluid immiscible with the sample fluid through the main channel, thereby forming droplets of the sample fluid in the extrusion fluid;

(b) flowing the droplets through the detection region;

(c) interrogating each droplet for a predetermined characteristic while the droplet flows through the detection region; and

(d) directing each droplet into a branched channel selected from (iv) according to results of step (c).

12. The method of claim 11 wherein the main channel has a diameter of between about 2 and 100 microns.

13. The method of claim 11, wherein the droplets formed in step (a) contain particles suspended in the sample fluid, and the characteristic interrogated in step (c) is a characteristic of at least some of the particles.

14. The method of claim 13 wherein the particle is a biological material.

15. The method of claim 14 wherein the biological material is a cell.

16. The method of claim 11, wherein the droplets formed in step (a) contain isolated DNA or protein.

17. The method of claim 11, wherein the main channel and the inlet channel are tapered towards the junction, and wherein the main channel and the inlet channel form a junction at an angle of about 60 to about 120 degrees.

18. The method of claim 11, wherein the sample fluid is introduced into the extrusion fluid as single monodisperse droplets with regular periodicity.

19. A method for analyzing droplets in a microfluidic device, comprising

(a) flowing a sample fluid containing particles through a first channel while flowing an extrusion fluid immiscible with the sample fluid through a second channel towards a junction of the two channels that is configured so that droplets of the sample fluid are introduced into the extrusion fluid;

(b) flowing the droplets formed in step (a) through a detection region that is downstream from the junction;

(c) interrogating each droplet while the droplet is flowing through the detection region; and

(d) assessing the presence or characteristics of a particle or material contained in each droplet from data obtained from the interrogating of step (c).

20. The method of claim 19, wherein step (c) comprises determining whether a reporter molecule is present in each droplet.