Method and device for isolating cells from heterogeneous solution using microfluidic trapping vortices

• US 9,133,499 B2
• Filed: 09/12/2011
• Issued: 09/15/2015
• Est. Priority Date: 09/14/2010
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Di Carlo, Dino, Hur, Soojung C., Mach, Albert J.
• Primary Examiner(s): Ware, Debbie K
• Assistant Examiner(s): Srivastava, Kailash C
• Application Number:
  US13/823,112
• Publication Number:
  US 20130171628A1
• Time in Patent Office: 1,464 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• 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 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/082 Active control of flow resistance, e.g. flow controllers
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• 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 3/502761 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 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 29/00 Means for introduction, extraction or recirculation of materials, e.g. pumps pumps per se F04B
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12Q 1/24 Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 1/4077 by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

A method of isolating cells includes providing a microfluidic device having at least one microfluidic channel coupled to an inlet and an outlet, the at least one microfluidic channel comprises at least one expansion region disposed along the length thereof. The at least one expansion region is an abrupt increase in a cross-sectional dimension of the at least one microfluidic channel configured to generate a vortex within the at least one expansion region in response to fluid flow. A solution containing a population of cells at least some of which have diameters ≧10 μm flows into the inlet. A portion of cells is trapped within vortex created within the at least one expansion region. The trapped cells may then released from the expansion region.

Claims

1. A method of isolating cells comprising:

providing a microfluidic device having at least one microfluidic channel coupled to an inlet and an outlet, the at least one microfluidic channel comprising a plurality of expansion regions disposed along the length thereof, each of the plurality of expansion regions comprising an abrupt increase in a cross-sectional dimension of the at least one microfluidic channel followed by an abrupt decrease in the cross-sectional dimension of the at least one microfluidic channel, wherein the plurality of expansion regions are configured to generate vortices within each expansion region in response to fluid flow;

flowing a solution containing a population of cells into the inlet;

trapping at least some of the cells within the vortices created within the plurality of expansion regions, the at least some of the cells having diameters ≧

10 μ

m; and

releasing the trapped cells from the plurality of expansion regions by reducing the flow rate of solution through the at least one microfluidic channel.

2. The method of claim 1, wherein the solution introduced into the inlet contains a homogenous population of cells.

3. The method of claim 1, wherein the solution introduced into the inlet contains a heterogeneous population of cells.

4. The method of claim 3, wherein the heterogeneous population of cells comprises a population containing cancer cells and wherein the cancer cells are trapped within the vortices.

5. The method of claim 1, wherein the at least one expansion region comprises a leading wall extending at least 45°

with respect to the axis of flow.

6. The method of claim 5, wherein the solution containing a heterogeneous population of cells comprises one of blood, urine, pleural fluid, and a peritoneal wash.

7. The method of claim 1, wherein the at least one expansion region has a profile of a rectangle, square, triangle, polygonal, or semi-circle.

8. The method of claim 1, wherein the released cells are collected at the outlet.

9. The method of claim 1, wherein cells having a diameter within the range of 10 μ

m to 30 μ

m are trapped within the vortex.

10. The method of claim 1, wherein the trapped cells comprise cancer cells.

11. The method of claim 1, wherein releasing the trapped cells comprises reducing the flow rate of solution through the at least one microfluidic channel to substantially zero and initiating the flow of a flushing solution.

12. The method of claim 1, wherein the microfluidic device comprises a plurality of microfluidic channels coupled to the inlet and the outlet.

13. The method of claim 12, wherein the plurality of microfluidic channels are arranged in a two-dimensional array.

14. The method of claim 12, wherein the plurality of microfluidic channels are arranged in a three-dimensional array.

15. The method of claim 1, wherein the abrupt increase in the cross-sectional dimension of the at least one microfluidic channel is at least 80 μ

m.

16. A method of isolating particles or cells by size comprising:

providing a microfluidic device having at least one microfluidic channel coupled to an inlet and an outlet, the at least one microfluidic channel comprising a plurality of expansion regions disposed along the length thereof, each of the plurality of expansion regions comprising an abrupt increase in a cross-sectional dimension of the at least one microfluidic channel followed by an abrupt decrease in the cross-sectional dimension of the at least one microfluidic channel, wherein the plurality of expansion regions are configured to generate vortices within the at least one expansion region in response to fluid flow;

flowing a solution containing a plurality of cells or particles into the inlet; and

trapping at least some of the cells or particles within the vortices created within the plurality of expansion regions, wherein the cells or particles having a size above a threshold value are substantially trapped within the vortices and wherein cells or particles having a size below the threshold value substantially pass by the vortices;

wherein the threshold value comprises a diameter of 10 μ

m.

17. The method of claim 16, further comprising releasing the trapped cells or particles from the plurality of expansion regions by reducing the flow rate of solution through the at least one microfluidic channel.

18. The method of claim 16, further comprising rapidly reducing the flow rate of solution through the at least one microfluidic channel to substantially zero.

19. The method of claim 16, wherein the plurality of expansion regions comprises a leading wall extending at least 45°

with respect to the axis of flow.

20. The method of claim 16, wherein the at least one expansion region has a profile of a rectangle, square, triangle, polygonal, or semi-circle.