Cell sorting apparatus and methods for manipulating cells using the same

• US 20040166555A1
• Filed: 02/13/2004
• Published: 08/26/2004
• Est. Priority Date: 11/10/1999
• Status: Abandoned Application

• Current Assignee: Joel Voldman, Martha Gray, Martin Schmidt, Mehmet Toner, Rebecca Braff
• Original Assignee: Joel Voldman, Martha Gray, Martin Schmidt, Mehmet Toner, Rebecca Braff
• Inventors: Braff, Rebecca, Schmidt, Martin, Toner, Mehmet, Voldman, Joel, Gray, Martha
• Application Number:
  US10/778,831
• Publication Number:
  US 20040166555A1
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00614 Delimitation of the attachment areas
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00704 integrated with the reactor apparatus
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: G01N 15/1456 without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
• CPC Class Codes: G01N 15/147 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 2015/1415 Control of particle position
• CPC Class Codes: G01N 30/0005 Field flow fractionation

Abstract

A cell analysis and sorting apparatus is capable of monitoring over time the behavior of each cell in a large population of cells. The cell analysis and sorting apparatus contains individually addressable cell locations. Each location is capable of capturing and holding a single cell, and selectively releasing that cell from that particular location. In one aspect of the invention, the cells are captured and held in wells, and released using vapor bubbles as a means of cell actuation. In another aspect of the invention, the cells are captured, held and released using electric field traps.

Claims

1. A method for analyzing a cell population, comprising:

providing a cell sorting apparatus having an array of sites, each site including a capture mechanism comprising a well that is capable of capturing a single cell, and a release mechanism comprising an actuator coupled to the well for selectively releasing the single cell from its site;

introducing a fluid medium containing a plurality of cells onto the apparatus;

capturing at least one cell in a well;

identifying a cellular response in the at least one captured cell; and

selecting at least one captured cell based on the cellular response.

2. The method of claim 1, wherein the step of capturing further comprises inducing a fluid medium with a cell population to flow across the substrate.

3. The method of claim 1, wherein the cell is captured in the well by negative pressure.

4. The method of claim 1, wherein the cell is captured in the well by well geometry.

5. The method of claim 1, wherein the cell is captured in the well by gravitational forces.

6. The method of claim 1, wherein the cell is captured in the well by electric field forces.

7. The method of claim 1, wherein the cell is captured in the well by dielectrophoretic forces.

8. The method of claim 1, wherein the step of identifying a cellular response further includes applying an optical probe to the captured cell and detecting an optical signal from the captured cell.

9. The method of claim 1, wherein a plurality of cells are captured, and the method further comprises the step of applying a fluorescent agent to the plurality of cells.

10. The method of claim 9, further including the step of detecting fluorescence exhibited by captured cells within the sorting apparatus.

11. The method of claim 10, further including the step of measuring intensity of fluorescence at individual sites in the sorting apparatus over time.

12. The method of claim 1, wherein the step of identifying a cellular response further comprises coupling a photometric array to the cell sorting apparatus to detect fluorescence at sites in the sorting apparatus.

13. The method of claim 12, further including the step of comparing the fluorescence intensity at particular sites.

14. The method of claim 13, wherein the step of selecting a cell further comprises selecting a cell based on its fluorescence intensity, and releasing the cell from its well.

15. The method of claim 13, wherein a receptor-ligand binding event at a site in the sorting apparatus is identified by fluorescence intensity.

16. The method of claim 13, wherein a protein to protein interaction at a site in the sorting apparatus is identified by fluorescence intensity.

17. The method of claim 13, wherein cellular signal transduction at a site in the sorting apparatus is identified by fluorescence intensity.

18. The method of claim 13, wherein intracellular calcium transport at a site in the sorting apparatus is identified by fluorescence intensity.

19. The method of claim 1, wherein the step of selecting at least one captured cell further comprises releasing the captured cell from its well by activating the release mechanism of the well.

20. The method of claim 19, wherein the step of releasing comprises forming a microbubble to displace the captured cell from its well.

21. The method of claim 19, wherein the step of releasing comprises lowering an energy field surrounding the captured cell.

22. A method for sorting individual cells in a cell population, comprising:

providing a cell sorting apparatus having an array of sites, each site including a capture mechanism comprising a well that is capable of capturing a single cell, and a release mechanism comprising an actuator coupled to the well for selectively releasing the single cell from its site;

introducing a fluid medium containing a plurality of cells onto the apparatus;

capturing at least one cell in a well;

selecting a captured cell; and

releasing the selected, captured cell from the apparatus.

23. The method of claim 22, wherein the step of capturing comprises inducing a fluid medium with a cell population to flow across the substrate.

24. The method of claim 22, wherein the cell is captured in the well by negative pressure.

25. The method of claim 22, wherein the cell is captured in the well by well geometry.

26. The method of claim 22, wherein the cell is captured in the well by gravitational forces.

27. The method of claim 22, wherein the cell is captured in the well by electric field forces.

28. The method of claim 22, wherein the cell is captured in the well by dielectrophoretic forces.

29. The method of claim 22, wherein a plurality of cells are captured within wells.

30. The method of claim 22, wherein the step of releasing the captured cell further comprises activating the actuator so as to release the captured cell from its well.

31. The method of claim 30, wherein the step of activating further comprises forming a microbubble to displace the captured cell from its well.

32. The method of claim 22, wherein the step of releasing the captured cell further comprises lowering an energy field surrounding the captured cell.

33. The method of claim 22, wherein the step of releasing the captured cell further comprises releasing the selected, captured cell into a fluid flow around the well.

34. The method of claim 22, further including the step of collecting at least one released cell.

35. The method of claim 22, further including the step of coupling a photometric array to the apparatus to detect fluorescence at sites in the sorting apparatus.

36. The method of claim 35, further including the step of applying a fluorescent agent to the captured cell.

37. The method of claim 36, further including the step of detecting fluorescence at sites within the sorting array.

38. The method of claim 37, wherein a temporal response of captured cells to stimuli is monitored.

39. The method of claim 22, wherein the step of selecting a captured cell from the sorting apparatus further comprises selecting the cell selected based on its response to at least one stimulus.

40. The method of claim 39, further including the step of observing a temporal response to stimulus.

41. The method of claim 39, wherein the captured cell is selected based on detection of a receptor-ligand binding event at a site in the sorting apparatus.

42. The method of claim 39, wherein the step of selecting the captured cell is based on detection of a protein to protein interaction at a site in the sorting apparatus.

43. The method of claim 39, wherein the captured cell is selected based detection of cellular signal induction at a site in the sorting apparatus.

44. The method of claim 39, wherein the captured cell is selected based on detection of intracellular calcium transport at a site in the sorting apparatus.

45. The method of claim 39, wherein the captured cell is selected based on detection of a drug response at a site in the sorting apparatus.

46. The method of claim 39, wherein the captured cell is selected based on detection of a phenotypical response at a site in the sorting apparatus.

47. The method of claim 46, wherein the phenotypical response of the selected cell is different from that of other cells captured in the sorting apparatus.

48. The method of claim 39, wherein the captured cell is selected based on a reporter-gene based assay.