Fractional cell sorter

• US 6,120,735 A
• Filed: 07/13/1999
• Issued: 09/19/2000
• Est. Priority Date: 02/26/1992
• Status: Expired due to Term

• Current Assignee: Cleveland Clinic Foundation, Ohio State Innovation Foundation (Ohio State University), Sony Corporation (Sony Group Corp.)
• Original Assignee: Cleveland Clinic Foundation, Ohio State University
• Inventors: Moore, Lee R., Zborowski, Maciej, Chalmers, Jeff
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): TRAN, HUAN T
• Application Number:
  US09/352,725
• Time in Patent Office: 434 Days
• Field of Search: 435/325, 435/173.4, 435/173.9, 435/2, 435/286.5, 422/73, 422/214, 422/216, 422/223.1, 422/231, 210/222, 210/223
• US Class Current: 422/73
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence

Abstract

A method and apparatus for the fraction sorting of cells is provided. The invention include a dispensing device, aspirating device, flow channel and a magnetic assembly. A first fractional sorting device includes a dipole fractional cell sorter. The dipole fractional cell sorter includes large aspect ratio flow channel and a dipole magnetic assembly. A second fractional sorting device includes a quadrupole fractional cell sorter. The quadrupole fractional cell sorter includes an annular flow channel having inlet and outlet splitters, and a quadrupole magnetic assembly. The quadrupole magnetic assembly includes pole pieces having projecting members with convex surfaces and in is in physical communication with the annular flow chamber. The method of the present invention provides a flow through fractional cell sorting process which is based on the application of a magnetic force to cells having a range of magnetic labeling densities.

Claims

1. A cell sorting device for the fractional sorting of cells, the device comprising:

(a) a dispensing device for providing a sample flow and a carrier flow;

(b) a flow channel for allowing the flow of the sample and the carrier;

(c) a first interface device for interfacing the dispensing device to the flow channel, the first interface device comprising an inlet injector region;

(d) an aspirating device for aspirating the flow present in the flow channel;

(e) a second interface device for interfacing the flow channel to the aspirating device, the second interface device comprising an exit port region; and

(f) a magnetic assembly for creating a magnetic field, the magnetic assembly comprising;

(i) a first and a second magnetic pole; and

(ii) an air gap disposed between the first and the second magnetic poles and configured to allow at least a partial insertion of the flow channel therein.

2. The device of claim 1 wherein the exit port region of the second interface device comprises a plurality of collectors in a parallel, configuration.

3. The device of claim 1 wherein the inlet injector region of the first interface device comprises a plurality of inlet injectors in a parallel, configuration.

4. The device of claim 2 wherein the plurality of collectors are equally spaced from each other.

5. The device of claim 3 wherein the plurality of inlet injectors are equally spaced from each other.

6. The device of claim 1 wherein the flow channel comprises a symmetrical cross-sectional geometry having a substantially uniform length and a substantially uniform width, wherein the length is substantially greater than the width.

7. The device of claim 6 wherein the flow channel is positioned within the magnetic field such that the vector direction of the magnetic forces created by the magnetic field are substantially parallel to the cross-sectional length of the flow channel.

8. The device of claim 5 wherein the plurality of inlet injectors comprises a sample injector that is disposed between carrier injectors.

9. The device of claim 1 wherein

the exit port region of the second interface device comprises a plurality of collectors,

the inlet injector region of the first interface device comprises a plurality of inlet injectors, and

the flow channel includes a plurality of flow compartments, and

wherein the plurality of collectors and the plurality of inlet injectors are further configured such that each collector and each inlet injector are within a flow compartment.

10. The device of claim 1 wherein

the exit port region of the second interface device comprises a plurality of collectors in a parallel, configuration,

the inlet injector region of the first interface device comprises a plurality of inlet injectors in a parallel, configuration, and

wherein the plurality of collectors and the plurality of inlet injectors are further configured such that each collector has a common center line with an inlet injector.

11. The device of claim 1 wherein the magnetic assembly is positioned substantially adjacent to the flow channel.

12. The device of claim 1 further comprising a fraction determining device for determining a sorted fraction distribution inside the flow channel.

13. The device of claim 3 wherein the plurality of inlet injectors comprises a sample injector which includes a conical-shaped discharge chamber having a large height-to-base diameter ratio.

14. The device of claim 3 wherein the plurality of inlet injectors comprises a plurality of carrier medium injectors and a sample injector and wherein the sample injector is substantially longer than the carrier medium injectors.

15. A cell sorting device for the fractional sorting of cells, the device comprising at least a first and a second stage,

(a) wherein the first stage comprises:

(1) a dispensing device for providing a sample flow and a carrier flow;

(2) a first stage flow channel for allowing the flow of the sample and the carrier;

(3) a first stage first interface device for interfacing the dispensing device to the flow channel, the first interface device comprising an inlet injector region;

(4) a first stage second interface device for interfacing the flow channel to the second stage, the second interface device comprising an exit port region; and

(5) a first stage magnetic assembly for creating a magnetic field, the magnetic assembly comprising;

a. a first and a second magnetic pole; and

b. an air gap disposed between the first and the second magnetic poles and configured to allow at least a partial insertion of the first stage flow channel therein; and

(b) wherein the second stage comprises;

(1) a second stage flow channel for allowing the flow of the sample and the carrier;

(2) a second stage first interface device for interfacing the first stage to the second stage, the first interface device comprising an inlet injector region;

(3) a second stage second interface device for interfacing the flow channel to an aspirating device for aspirating the flow present in the flow channel, the second interface device comprising an exit port region; and

(4) a second stage magnetic assembly for creating a magnetic field, the magnetic assembly comprising;

a. a first and a second magnetic pole; and

b. an air gap disposed between the first and the second magnetic poles and configured to allow at least a partial insertion of the second stage flow channel therein.

16. The device of claim 15 wherein the flow channel of the first and second stages comprises a symmetrical cross-sectional geometry having a substantially uniform length and a substantially uniform width, wherein the length is substantially greater than the width.

17. The device of claim 16 wherein the flow channel of the first and second stages is positioned within the magnetic field of each stage such that the vector direction of the magnetic forces created by the magnetic field of each stage is substantially parallel to the cross-sectional length of the flow channel of each stage.

18. The device of claim 15 further comprising a fraction determining device for determining a sorted fraction distribution.

19. A cell sorting device for the fractional sorting of cells, the device comprising:

(a) a dispensing device for providing a sample flow and a carrier flow;

(b) an annular flow channel for allowing the flow of the sample and the carrier;

the annular flow channel having an inlet and an outlet;

(c) a first annular splitter disposed at least partially within the inlet of the annular flow channel;

(d) a second annular splitter disposed at least partially within the outlet of the annular flow channel;

(e) an aspirating device for aspirating the flow present in the annular flow channel; and

(f) a magnetic assembly for creating a magnetic field, the magnetic assembly comprising;

(i) first, second, third and forth magnetic poles; and

(ii) an air gap disposed between the first, second, third and forth magnetic poles and configured to allow at least a partial insertion of the annular flow channel therein.

20. The device of claim 19 wherein each magnetic pole comprises a projection member having a convex surface.

21. The device of claim 20 wherein the annular flow channel further comprises an outer and inner cylindrical wall and wherein the convex surface of each magnetic pole is in physical communication with the annular flow channel outer wall.

22. The device of claim 19 wherein the first, second, third and forth magnetic poles each comprise a permanent magnet and a pole piece.

23. The device of claim 21 wherein the air gap comprises a magnetic field having a magnetic force that includes a vector direction which is radially outward towards the annular flow channel outer wall.

24. The device of claim 19 wherein the annular flow channel further comprises:

(a) an outer wall;

(b) an inner wall; and

(c) an annular separation region disposed between the inner and outer walls for allowing the migration of magnetically labeled cells.

25. A cell sorting device for the fractional sorting of cells, the device comprising:

(a) a dispensing device for providing a sample flow and a carrier flow;

(b) an aspirating device for aspirating the flow present in the annular flow channel;

(c) at least a first and a second sorting stage for sorting a predetermined fraction of cells having a predetermined range of densities of a particular cell characteristic, wherein the first and second sorting stages each comprise;

(1) an annular flow channel for allowing the flow of the sample and the carrier;

the annular flow channel having an inlet and an outlet;

(2) a first annular splitter disposed at least partially within the inlet of the annular flow channel;

(3) a second annular splitter disposed at least partially within the outlet of the annular flow channel; and

(4) a magnetic assembly for creating a magnetic field, the magnetic assembly comprising;

(i) first, second, third and forth magnetic poles; and

(ii) an air gap disposed between the first, second, third and forth magnetic poles and configured to allow at least a partial insertion of the annular flow channel therein.

26. The device of claim 25 wherein the outlet of the annular flow channel of the first sorting stage comprises a first fraction outlet port and wherein the inlet port of the annular flow channel of the second stage comprises a sample injection port and wherein the first fraction outlet port of the first sorting stage is in fluid communication with the sample injection port of the second sorting stage.

27. The device of claim 25 wherein each magnetic pole comprises a projection member having a convex surface.

28. The device of claim 25 wherein each annular flow channel further comprises:

(a) an outer wall;

(b) an inner wall; and

(c) an annular separation region disposed between the inner and outer walls for allowing the migration of magnetically labeled cells.