Multiaperture sample positioning and analysis system

• US 20030146091A1
• Filed: 12/31/2002
• Published: 08/07/2003
• Est. Priority Date: 12/17/1997
• Status: Active Grant

• Current Assignee: Molecular Devices, Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Original Assignee: MDS Analytical Technologies US Incorporated (Danaher Corporation)
• Inventors: Schmidt, Christian, Vogel, Horst
• Granted Patent: US 7,387,715 B2
• Time in Patent Office: Days
• US Class Current: 204/403.01
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: G01N 33/48728 Investigating individual cells, e.g. by patch clamp, voltage clamp investigating individual particles in general G01N15/10

Abstract

Systems for positioning and/or analyzing samples such as cells, vesicles, cellular organelles, and fragments, derivatives, and mixtures thereof, for electrical and/or optical analysis, especially relating to the presence and/or activity of ion channels.

Claims

1. A system for analyzing membranous samples, comprising:

a carrier with a shelf structure, the shelf structure having an aperture;

first and second fluid compartments, separated by the carrier, and fluidically connected to one another, in the absence of a membranous sample, via the aperture;

a membranous sample, selected from the group consisting of cells, organelles, and vesicles, sealed across the aperture, wherein the seal has an electrical resistance of greater than about 1 GΩ

; and

first and second electrodes, in electrical contact with the first and second fluid compartments, respectively, wherein the first and second electrodes are adapted to apply and/or measure at least one of an electrical potential and a current from at least a portion of the membranous sample sealed across the aperture.

2. The system of claim 1, wherein the sample is a cell.

3. The system of claim 1, wherein the carrier is at least substantially planar.

4. The system of claim 1, wherein the carrier is formed at least in part of silicon.

5. The system of claim 1, wherein the carrier includes first and second layers having different compositions.

6. The system of claim 5, wherein the shelf structure is associated with a transition between the first and second layers.

7. The system of claim 1, wherein the shelf structure is associated with an abrupt change in the cross-sectional area of the aperture.

8. The system of claim 1, wherein the shelf structure is associated with a change in the composition of the carrier.

9. The system of claim 1, wherein the shelf structure is positioned adjacent an end of the aperture.

10. The system of claim 1, wherein the shelf structure has first and second sides facing the first and second fluid compartments, respectively, and wherein at least one of the sides is at least substantially planar.

11. The system of claim 1, wherein the shelf structure has first and second sides facing the first and second fluid compartments, respectively, and wherein at least one of the sides is bent.

12. The system of claim 1, wherein the aperture is characterized by an aperture axis that extends between opposite ends of the aperture, and wherein the shelf structure includes at least one surface that is at least substantially perpendicular to the aperture axis.

13. The system of claim 1, wherein the shelf structure comprises a ledge.

14. The system of claim 1, wherein at least a portion of the shelf structure is annular in shape.

15. The system of claim 1, wherein a surface of the shelf structure is modified to promote sample adhesion.

16. The system of claim 15, wherein the surface is electrically charged.

17. The system of claim 15, wherein the surface is hydrophilic.

18. The system of claim 15, wherein the surface is plasma cleaned before the carrier is modified or immediately before the carrier is used.

19. The system of claim 1, wherein the carrier includes a plurality of apertures, each aperture located on a shelf structure.

20. The system of claim 1, wherein the membranous sample is supported by the shelf structure.

21. The system of claim 20, wherein the membranous sample is positioned at least substantially outside the aperture.

22. The system of claim 20, wherein the membranous sample is positioned at least substantially inside the aperture.

23. The system of claim 1, wherein the aperture is formed at least in part by anisotropic etching.

24. The system of claim 1 further comprising a sample-analysis mechanism adapted to perform a fluorescence measurement on a sample positioned about the aperture.

25. The system of claim 1 further comprising a sample-handling mechanism configured to add at least one of a sample and a fluid to at least one of the fluid compartments.

26. The system of claim 25, wherein the sample-handling mechanism is configured to introduce samples into at least one compartment through an opening arranged above the aperture.

27. The system of claim 25, wherein the sample-handling mechanism is configured to check the fluid level in at least one compartment.

28. A system for analyzing membranous samples, comprising:

a carrier having an opening, the interior of the opening including a shelf structure;

first and second fluid compartments, separated by the carrier, and fluidically connected to one another, in the absence of a membrane sample, via the opening;

first and second electrodes, in electrical contact with the first and second fluid compartments, respectively, wherein the first and second electrodes being adapted to apply and/or measure at least one of an electrical potential and a current from at least a portion of a membranous sample sealed within the opening; and

a membranous sample, selected from the group consisting of cells, organelles, and vesicles, sealing the opening.

29. The system of claim 28, wherein the sample seals the opening with an electrically tight bond.

30. The system of claim 28, wherein the opening has a wall surrounding an interior, wherein the shelf structure projects inwardly from the wall toward the interior.

31. The system of claim 28, wherein the interior of the opening further includes a funnel structure that tapers toward or away from the shelf structure.

32. The system of claim 28, wherein the opening includes the first or second or both fluid compartments.

33. A system for analyzing membranous samples, comprising:

a carrier having a planar surface and an aperture terminating at the planar surface;

first and second fluid compartments, separated by the carrier, and fluidically connected, in the absence of a sample, via the aperture;

a membranous sample, selected from the group consisting of cells, organelles, and vesicles, sealed across the aperture, wherein the seal has an electrical resistance of greater than about 1 gigaohm; and

first and second electrodes, in electrical contact with the first and second fluid compartments, respectively, wherein the first and second electrodes are adapted to apply and/or measure at least one of an electrical potential and a current from at least a portion of the membranous sample sealed across the aperture.

34. The system of claim 33, wherein the membranous sample is a cell.

35. The system of claim 33, wherein the planar surface is a first planar surface, and wherein the carrier includes a second planar surface, at least generally parallel to the first planar surface.

36. The system of claim 35, wherein the aperture has two ends, and wherein a first end terminates at the first planar surface, and a second end terminates at least generally at the second surface.

37. The system of claim 35, wherein the aperture is characterized by an aperture axis that extends between opposite ends of the aperture, and wherein the aperture axis is at least substantially perpendicular to the first and second surfaces.

38. The system of claim 33, wherein the carrier has a shelf structure, and wherein the aperture traverses the shelf structure.

39. The system of claim 33, wherein the carrier includes first and second layers having different compositions, and wherein the aperture traverses the first and second layers.

40. The system of claim 33, wherein a surface of at least one of the layers is modified to promote sample adhesion.

41. The system of claim 33, wherein the carrier includes a plurality of apertures, each aperture disposed in a planar surface.

42. The system of claim 33 further comprising a sample-analysis mechanism adapted to perform a fluorescence measurement on a sample positioned about the aperture.

43. The system of claim 33 further comprising a sample-handling mechanism configured to add at least one of a sample and a fluid to at least one of the fluid compartments.

44. A system for analyzing membranous samples, comprising:

a carrier comprising (i) a plurality of layers, the plurality including at least a first layer and a second layer having different compositions, and (ii) an aperture that traverses the first and second layers;

first and second fluid compartments, separated by the carrier, and fluidically connected to one another, in the absence of a sample, via the aperture;

a membranous sample, selected from the group consisting of cells, organelles, and vesicles, sealed across the aperture; and

first and second electrodes, in electrical contact with the first and second fluid compartments, respectively, wherein the first and second electrodes are adapted to apply and/or measure at least one of an electrical potential and a current from at least a portion of the membranous sample sealed across the aperture.

45. The system of claim 44, wherein the carrier is at least substantially planar.

46. The system of claim 45, wherein the aperture spans the space between opposed surfaces of the carrier, and wherein, adjacent the aperture, the first and second layers are at least substantially parallel to the opposed surfaces.

47. The system of claim 45, wherein the aperture is characterized by an aperture axis that extends between opposite ends of the aperture, and wherein the aperture axis is at least substantially perpendicular to the plane of the carrier.

48. The system of claim 44, wherein the seal between the sample and the aperture has an electrical resistance of at least about 1 gigaohm.

49. The system of claim 44, wherein at least one of the first and second layers includes silicon.

50. The system of claim 44, wherein the aperture is at least substantially tapered in the first layer and at least substantially cylindrical in the second layer.

51. The system of claim 44, wherein the average diameter of the aperture in at least one of the first and second layers is comparable to the average thickness of the layer.

52. The system of claim 44, wherein the first and second layers have different thicknesses.

53. The system of claim 44, wherein the first and second layers have different etchabilities.

54. The system of claim 44, wherein a surface of at least one of the layers is modified to promote sample adhesion.

55. The system of claim 44, wherein the carrier includes a plurality of apertures, each of the apertures traversing the first and second layers.

56. The system of claim 44 further comprising a sample-analysis mechanism adapted to perform a fluorescence measurement on a sample positioned about the aperture.

57. The system of claim 44 further comprising a sample-handling mechanism configured to add at least one of a sample and a fluid to at least one of the fluid compartments.

58. A system for analyzing membranous samples, comprising:

a carrier comprising (i) an aperture, and (ii) a binding surface associated with the aperture, the binding surface being adapted to bind a membranous sample such that the sample seals across the aperture;

first and second fluid compartments, separated by the carrier, and fluidically connected to one another, in the absence of a sample, via the aperture;

a membranous sample, selected from the group consisting of cells, organelles, and vesicles, bound to the binding surface, and sealed across the aperture; and

first and second electrodes, in electrical contact with the first and second fluid compartments, respectively, wherein the first and second electrodes are adapted to apply and/or measure at least one of an electrical potential and a current from at least a portion of the membranous sample sealed across the aperture.

59. The system of claim 58, wherein the carrier is at least substantially planar.

60. The system of claim 58, wherein the binding surface is adjacent the aperture.

61. The system of claim 58, wherein the binding surface is within the aperture.

62. The system of claim 58, wherein the seal between the sample and the aperture has an electrical resistance of at least about 1 gigaohm.

63. The system of claim 58, wherein the binding surface is electrically charged, such that it attracts biological membranes.

64. The system of claim 63, wherein the electrical charge is produced with the aid of polycations and/or silanes.

65. The system of claim 58, wherein the binding surface provides a molecule-specific bonding of cells, vesicles, membrane fragments, or biological organelles.

66. The system of claim 65, wherein the molecule-specific bonding is mediated by biotin-streptavidin interactions or histidine-nitriloacetic acid interactions.

67. The system of claim 58, wherein the binding surface provides a multivalent-ion-mediated bonding of cells, vesicles, membrane fragments, or biological organelles.

68. The system of claim 67, wherein the multivalent-ion-mediated bonding is mediated by Ca²⁺ ions.

69. The system of claim 58, wherein the binding surface has hydrophilic and hydrophobic regions, and wherein the hydrophilic regions are disposed around the apertures.

70. The system of claim 58, wherein the carrier is plasma cleaned before the surface of the carrier is modified or immediately before the carrier is used.

71. The system of claim 58, wherein the sample is modified to enhance bonding between the sample and the carrier.

72. The system of claim 71, wherein the binding surface is charged, and wherein the sample is modified by charging its surface with a charge opposing that on the binding surface.

73. The system of claim 58, wherein the carrier includes a plurality of apertures, each of the apertures being adjacent a binding surface adapted to bind a sample such that the sample seals across the corresponding aperture.

74. The system of claim 58 further comprising a sample-analysis mechanism adapted to perform a fluorescence measurement on a sample positioned about the aperture.

75. The system of claim 58 further comprising a sample-handling mechanism configured to add at least one of a sample and a fluid to at least one of the fluid compartments.