Methods and apparatus for sorting cells using an optical switch in a microfluidic channel network

• US 8,426,209 B2
• Filed: 06/28/2010
• Issued: 04/23/2013
• Est. Priority Date: 08/28/2003
• Status: Active Grant

• Current Assignee: Progenity, Inc. (Athyrium Capital Management LP)
• Original Assignee: Celula, Inc. (Neoseq Ltd.)
• Inventors: Butler, William Frank, Chachisvilis, Mirianas, Dees, Robert, Hagen, Norbert, Marchand, Philippe, Raymond, Daniel E., Tu, Eugene, Wang, Mark M., Yang, Joon Mo, Yang, Rong, Zhang, Haichuan
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): Handy, Dwayne K
• Application Number:
  US12/825,037
• Publication Number:
  US 20100304429A1
• Time in Patent Office: 1,030 Days
• Field of Search: 436/63, 436/164, 436/165, 436/172, 436/174, 436/180
• 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 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 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0454 radiation pressure, optical tweezers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• 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 3/502761 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 3/502776 specially adapted for focusing or laminating flows
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

Apparatus and Methods are provided for a microfabricated fluorescence activated cell sorter based on an optical switch for rapid, active control of cell routing through a microfluidic channel network. This sorter enables low-stress, highly efficient sorting of populations of small numbers of cells (i.e., 1000-100,000 cells). The invention includes packaging of the microfluidic channel network in a self-contained plastic cartridge that enables microfluidic channel network to macro-scale instrument interconnect, in a sterile, disposable format.

Claims

1. A method for cell sorting in a device comprising a microfluidic network having a sample inlet, one or more inlet channels, a main channel, and at least two outlet channels, comprising the steps of:

receiving cells in a fluidic medium at the sample inlet,

flowing the cells through the main channel, the main channel being divided into one side and another side as defined by the path of the flow of cells,

subjecting the cells to a bias flow preferentially resulting in collection of the cells in a first reservoir in fluid communication with a first outlet channel, wherein the bias flow is created by applying asymmetric pressure to one or more of the channels of the microfluidic network,

identifying a cell to be sorted through application of a lateral force into a second reservoir coupled to a second outlet channel, and

applying a lateral force on the cell, the lateral force consisting of a single force applied from one side of the main channel and not from the said another side of the main channel, and characterized in that the lateral force is a non-trapping force, whereby the cell is moved transverse to the preferentially biased flow and selectively exits the second outlet channel coupled to the second reservoir.

2. The method for cell sorting of claim 1, wherein subjecting the cell to a bias flow by applying asymmetric pressure comprises introducing a pressure differential across the at least two outlet channels to direct the cell toward the first outlet channel coupled to the first reservoir.

3. The method for cell sorting of claim 2, further comprising separately controlling the pressure at the at least two outlet channels to create the pressure differential, wherein the pressure at each of the at least two outlet channels is selected from the group consisting of:

above atmospheric pressure, atmospheric pressure, and below atmospheric pressure.

4. The method for cell sorting of claim 1, wherein subjecting the cell to a bias flow by applying asymmetric pressure comprises introducing a pressure differential by using at least one pneumatic controller to separately control pressure at one or more of the outlet channels, the sample inlet channel and the inlet channels.

5. The method for cell sorting of claim 1, further comprising at least two inlet channels wherein subjecting the cell to a bias flow by applying asymmetric pressure comprises introducing a pressure differential across the at least two inlet channels to direct the cell toward the first outlet channel coupled to the first reservoir.

6. The method for cell sorting of claim 5, further comprising separately controlling the pressure at the at least two inlet channels to create the pressure differential, wherein the pressure at each of the at least two inlet channels is selected from the group consisting of:

above atmospheric pressure, atmospheric pressure, and below atmospheric pressure.

7. The method for cell sorting of claim 1, wherein the first reservoir is a non-target cell reservoir.

8. The method for cell sorting of claim 1, wherein the second reservoir is a target cell reservoir.

9. A method for cell sorting in a device comprising a microfluidic network having a sample inlet, one or more inlet channels, a main channel, and at least two outlet channels, comprising the steps of:

receiving cells in a fluidic medium at the sample inlet,

flowing the cells through the main channel,

subjecting the cells to a bias flow preferentially resulting in collection of the cells in a first reservoir in fluid communication with a first outlet channel, wherein the bias flow is created by configuring one or more of the channels of the microfluidic network to have asymmetric geometries,

identifying a cell to be sorted through application of a lateral force into a second reservoir coupled to a second outlet channel, and

applying a lateral force on the cell, consisting of a single lateral force configured to be translated at a non-zero angle down the main channel toward the at least two outlet channels and applied from one side of the main channel, and characterized in that the lateral force is a non-trapping force, whereby the cell is moved transverse to the preferentially biased flow and selectively exits the second outlet channel coupled to the second reservoir.

10. The method of cell sorting of claim 9, wherein the bias flow is created by configuring the at least two outlet channels to have asymmetric geometries.

11. The method of cell sorting of claim 10, wherein creating the bias flow by configuring the at least two outlet channels to have asymmetric geometries comprises configuring the first outlet channel to have a larger volumetric flow relative to the second outlet channel.

12. The method of cell sorting of claim 11, wherein configuring the first outlet channel to have a larger volumetric flow relative to the second outlet channel comprises configuring the first outlet channel to have a larger channel width relative to the second outlet channel.

13. The method of cell sorting of claim 9, further comprising at least two fluid inlet channels wherein the bias flow is created by configuring the first and second inlet channels to have asymmetric geometries.

14. The method of cell sorting of claim 13, wherein creating the bias flow by configuring the first and second inlet channels to have asymmetric geometries comprises configuring a first fluid inlet channel to have a larger volumetric flow relative to a second fluid inlet channel.

15. The method of cell sorting of claim 14, wherein configuring the first inlet channel to have a larger volumetric flow relative to the second inlet channel comprises configuring the first inlet channel to have a larger channel width relative to the second inlet channel.

16. The method of cell sorting of claim 9, wherein the microfluidic network further comprises at least two fluid inlet channels and wherein subjecting the cell to a bias flow by configuring one or more of the channels of the microfluidic network to have asymmetric geometries comprises configuring inlet channels to provide equal flow rates and configuring the first outlet channel to have a larger volumetric flow relative to the second outlet channel.

17. The method of cell sorting of claim 9, wherein the microfluidic network further comprises at least two fluid inlet channels and wherein subjecting the cell to a bias flow by configuring one or more of the channels of the microfluidic network to have asymmetric geometries comprises configuring the inlet channels to have symmetric geometries and configuring the outlet channels to have asymmetric geometries.

18. The method of cell sorting of claim 9, wherein the first reservoir is a non-target cell reservoir.

19. The method of cell sorting of claim 9, wherein the second reservoir is a target cell reservoir.

20. The method for cell sorting of claim 9, wherein identifying a cell to be sorted comprises using at least one detection modality selected from the group comprising fluorescent and non-fluorescent detection modalities.

21. A method for cell sorting in a device comprising a microfluidic network having a sample inlet, one or more inlet channels, a main channel, and at least two outlet channels, comprising the steps of:

receiving cells in a fluidic medium at the sample inlet,

flowing the cells through the main channel, the main channel being divided into one side and another side as defined by the path of the flow of cells,

subjecting the cells to a bias flow preferentially resulting in collection of the cells in a first reservoir in fluid communication with a first outlet channel, wherein the bias flow is created by applying asymmetric pressure to one or more of the channels of the microfluidic network,

identifying a cell to be sorted through application of a lateral force into a second reservoir coupled to a second outlet channel, and

applying a lateral force on the cell, the lateral force consisting of a single force applied from one side of the main channel and characterized in that the lateral force is a non-trapping force, whereby the cell is moved transverse to the preferentially biased flow and selectively exits the second outlet channel coupled to the second reservoir.