FLAT CELL CARRIERS WITH CELL TRAPS

• US 20110117634A1
• Filed: 04/21/2009
• Published: 05/19/2011
• Est. Priority Date: 04/21/2008
• Status: Abandoned Application

• Current Assignee: Asaf Halamish, Lilach Weisz, Michael Sister
• Original Assignee: Asaf Halamish, Lilach Weisz, Michael Sister
• Inventors: Weisz, Lilach, Halamish, Asaf, Sister, Michael
• Application Number:
  US12/988,806
• Publication Number:
  US 20110117634A1
• Time in Patent Office: Days
• US Class Current: 435/283.1
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093

Abstract

Cell carrier structures using dynamic flow of cell bearing, washing, or nourishing fluid, from an input reservoir region to an output reservoir region. A common feature of the several structures by which this can be achieved is the presence of channels generally having low height or other cross section, such that the cell bearing fluid readily traverses these channels by capillary action. Optional pumping assistance can also be provided. The capture wells or traps are disposed generally along the length of these channels such that the cells have multiple chances of being captured in a trap or well. The traps or wells are structured such that only a single cell can be trapped in each well or trap, and the disposition of the wells or traps as appendages to the fluid flow channels facilitates the washing or nourishing of the cells while their proliferation or development is being observed.

Claims

1. A cell carrier for capturing cells, comprising:

a planar body member in which is formed an array of cell capturing wells, said cell capturing wells generally comprising an entrance aperture open to one surface of said body member and a plurality of openings at the end of said well distant from said entrance aperture and disposed at least partly in the well wall, such that not all of said openings can be simultaneously blocked by the presence of a captured cell, said entrance aperture having dimensions relative to that of the cells to be captured, such that only a single cell at a time can enter a well;

a fluid collection passage in fluid communication with said plurality of openings of said cell capturing wells; and

a pumping port in fluid communication said fluid collection passage,

wherein said openings have cross sectional dimensions significantly smaller than those of said entrance apertures.

2. A cell carrier according to claim 1 wherein said openings are such that a cell of size that said cell capturing well is adapted to capture, cannot pass through said openings.

4. A cell carrier according to claim 1, wherein said plurality of openings at the end of said well are disposed off the axis of said well, such that not all of said openings can be simultaneously blocked by the presence of a captured cell.

5. A cell carrier according to claim 1, wherein the height of said fluid collection passage is sufficiently small that fluid disposed in said pumping port flows through said fluid collection passage by capillary action.

6. A cell carrier according to claim 5, wherein said plurality of openings enables fluid flowing through said fluid collection passage to rise into said cell capture wells.

7. A cell carrier according to claim 1, further comprising a fluid application region in fluid contact with said one surface of said body member, such that fluid deposited in said fluid application region accesses said entrance apertures of said capture wells.

8. A cell carrier according to claim 7, further comprising a cover positioned on said cell carrier covering said fluid application area and said one surface of said body member, such that fluid applied to said fluid application area flows by capillary action to said one surface of said body member.

14. A cell carrier according to claim 1, wherein application of pumping action to said pumping port is operative to hold a captured cell at said at least one opening at the bottom end of said at least one cell capture chamber, such that a second cell of similar size cannot enter the entrance aperture into said well.

15. A cell carrier according to claim 14, wherein said cell capturing well is widened at its end distant from said entrance aperture, and release of pumping action from said pumping port is operative to release said captured cell such that it can spread laterally within said widened region.

9-13. -13. (canceled)

16. A cell carrier, comprising:

a base from which a plurality of rows of walls protrude;

a cover disposed in contact with at least some of the ends of said walls distant from said base, such that at least one closed flow channel is formed between an adjacent pair of walls, said base and said cover; and

at least one inlet in fluid connection with one end of said at least one flow channel and at least one outlet in fluid connection with a second end of said at least one flow channel, such that a fluid applied at said at least one inlet flows along said at least one flow channel to said at least one outlet,

wherein said at least one flow channel has a plurality of protrusions positioned down its length, such that cell capture traps are formed between said protrusions and said walls.

17. A cell carrier according to claim 16 and wherein said plurality of protrusions comprises either lateral protrusions attached to said walls along their length, or protrusions extending from at least one of said base and said cover, positioned close to said walls along their length.

21. A cell carrier according to claim 16 and wherein at least some of said cell traps along the length of said at least one flow channel have entrance openings aligned to face into the direction from which said fluid flows.

22. A cell carrier according to claim 21 and wherein at least some of said cell traps have outflow openings at the end opposite to said entrance openings, said outflow openings being smaller in cross section than said entrance openings, such that a cell directed into a cell trap cannot pass through its outflow opening.

23. A cell carrier according to claim 21, wherein said outflow openings allow a flow of fluid from said at least one flow channel through said at least some cell traps, such that cells borne by said fluid flow are directed into said cell traps.

27. A cell carrier according to claim 16, wherein at least some of said protrusions are disposed down said at least one flow channel at locations opposite the entrances of cell traps on the opposite side of said at least one flow channel, such that said lateral protrusions encourage entry of cells into said cell traps on the opposite side of said at least one flow channel.

28. A cell carrier according to claim 16, wherein said protrusions are positioned such as to generate zig-zag motion of fluid down said at least one flow channel, such that cells having a higher density than said fluid are directed into said traps, while said fluid continues its zig-zag motion down said at least one flow channel.

18-20. -20. (canceled)

24-26. -26. (canceled)

29. A cell carrier, comprising:

a base plate;

a cover plate; and

a cell trapping structure disposed between said base plate and said cover plate, said cell trapping structure comprising;

a plurality of sets of double walls, each set of double walls defining a first channel between them, and the spaces between neighboring sets of double walls defining a second channel, at least some of said double walls having protrusions disposed along their length on those sides of said walls that project into said second channel, such that the regions between adjacent protrusions constitute cell traps;

wherein said cover plate and said base plate contact at least some of said walls such that closed flow channels are formed therebetween, said cover plate comprising at least a first port in fluid connection with a reservoir at one end of at least some of said second channels; and

at least a second port in fluid connection with a reservoir at a second end of at least some of said second channels, and at least a third port in fluid connection with one end of at least some of said first channels, the other ends of which are sealed,

and wherein at least some of said cell traps have orifices at their wall ends, said orifices providing fluid contact between said cell traps and said first channels.

30. A cell carrier according to claim 29, wherein the application of suction to said third port generates an accompanying suction effect in said cell traps, directing fluid flowing in at least some of said second channels into at least some of said cell traps, such that at least some cells borne in said fluid flowing in said at least some second channels are trapped in some of said cell traps.

33. A cell carrier according to claim 29, wherein said at least a first port in said cover plate is operative to input fluid to said second channels and said at least second port in said cover plate is operative to output fluid from said second channels.

35. A cell carrier according to claim 29, wherein said orifices have dimensions such that a cell directed into a cell trap and having dimensions such that only a single such cell can enter said cell trap, cannot pass through said orifice.

36. A cell carrier according to claim 29, wherein said cell trapping structure is constructed as an integral part of either one of said cover plate and said base plate.

37. A cell carrier according to claim 29, wherein at least one of said cover plate and said base plate is constructed of a flexible material such that at least one of them can, when said cell carrier is under positive pressure, separate from contact with said cell trapping structure, such that said fluid can flow more readily into said flow channels.

31-32. -32. (canceled)

34. (canceled)