Multiaperture sample positioning and analysis system

• US 7,201,836 B2
• Filed: 03/07/2002
• Issued: 04/10/2007
• Est. Priority Date: 12/17/1997
• Status: Expired due to Term

• Current Assignee: Molecular Devices, Ecole polytechnique fãdãrale de lausanne epfl (Schweizerische Eidgenossenschaft)
• Original Assignee: Molecular Devices
• Inventors: Schmidt, Christian, Vogel, Horst
• Primary Examiner(s): Olsen; Kaj K.
• Application Number:
  US10/093,680
• Publication Number:
  US 20030052002A1
• Time in Patent Office: 1,860 Days
• Field of Search: 204/403.01, 204/403.03, 204/416, 205/777.5, 205/778, 435/4, 435/173.4, 435/173.6, 435/287.1, 435/288.4, 435/288.7
• US Class Current: 205/777.5
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/5088 confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• 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 multiaperture system for analyzing membranous samples, comprising:

a carrier having a plurality of apertures;

at least two fluid compartments, separated by the carrier, and fluidically connected to one another, in the absence of a membranous sample, via at least one of the apertures, at least one fluid compartment including a suspension, and a substantially planar seal surface circling the aperture;

a membranous sample from the suspension, selected from the group consisting of cells, organelles, and vesicles, sealed across one of the plurality of apertures, wherein the seal between the sample and the carrier is tight enough so that the variance of leakage currents through the aperture during electrical measurements lies below the signals to be measured by a factor of 5–

10; and

at least two electrodes in electrical contact with at least one of the fluid compartments, wherein the electrodes are adapted to apply and/or measure at least one of an electrical potential and a current, across at least a portion of the membranous sample sealed across the one of the plurality of apertures within the seal surface.

2. The system of claim 1, wherein the seal has an electrical resistance of greater than about 1 GΩ

.

3. The system of claim 1, wherein the variance of leakage currents tics below the signals lobe measured by a factor of 5–

10 when the signals to be measured lie between about 0.1 pA and about 50 pA at V_M=−

60 mV.

4. The system of claim 1, wherein each fluid compartment is in electrical contact with at least one electrode.

5. The system of claim 1, wherein the membranous sample is selected from the group consisting of cells, vesicles, and biological organelles.

6. The system of claim 1, wherein at least one electrode is used for measurement over more than one aperture.

7. The system of claim 1, the carrier having first and second sides, wherein several or all of the electrodes on one of the first and second sides of the carrier have a common electrical potential.

8. The system of claim 1, the carrier having first and second sides, wherein the compartments and electrodes are separated on one of the first and second sides of the carrier to allow independent measurements.

9. The system of claim 1, wherein the system is configured for sequential measurements over at least two apertures.

10. The system of claim 1, wherein the system is configured for parallel measurements over at least two apertures.

11. The system of claim 1, further comprising a sample-handling mechanism for adding, manipulating, exchanging, storing, and/or removing samples and sample components from at least one compartment.

12. The system of claim 11, the carrier having first and second sides, wherein the sample-handling mechanism is configured to add and/or store fluid on one or both of the first and second sides of the carrier.

13. The system of claim 11, the carrier having first and second sides, wherein the sample-handling mechanism is configured to add cells, vesicles, and other biological organelles or particles on one or both of the first and second sides of the carrier.

14. The system of claim 11, wherein the sample-handling mechanism is configured to check the fluid level in the compartments.

15. The system of claim 11, wherein the sample-handling system is configured to introduce samples into a compartment through an inlet opening arranged concentrically above the aperture.

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

17. The system of claim 1, wherein the carrier has an electrically charged surface that attracts biological membranes.

18. The system of claim 1, wherein the carrier has a surface that provides a molecule-specific bonding of cells, vesicles, or biological organelles on it.

19. The system of claim 1, wherein the carrier is formed at least in part of materials selected from the group consisting of silicon, silicon derivatives, glass, and plastic.

20. The system of claim 1, wherein the carrier comprises a silicon chip or a silicon wafer.

21. The system of claim 1, the carrier having first and second sides, wherein the compartments on at least one of the first and second sides of the carrier are open.

22. The system of claim 1, wherein at least two fluid compartments are disposed on one side of the carrier, each of those compartments being in contact, via the apertures, with a single fluid compartment disposed on the other side of the carrier.

23. The system of claim 1, further comprising an optical-analysis mechanism configured for optical analysis of samples positioned about at least one aperture.

24. The system of claim 23, wherein the optical-analysis mechanism is configured for fluorescence measurements.

25. The system of claim 1, further comprising a holder adapted to support the carrier.

26. The system of claim 1, wherein the system includes a row of 8, 12, 16, 24, 32, or 48 apertures.

27. The system of claim 1, wherein the application of a voltage difference, mediated by the electrodes, creates an inhomogeneous electric field around the aperture, which becomes larger as the aperture is approached.

28. The system of claim 1, the membranous sample being a first membranous sample, further comprising a second membranous sample, selected from the group consisting of cells, organelles, and vesicles, and fragments, derivatives, and mixtures thereof, scaled across a second one of the plurality of apertures, wherein the seal between the second membranous sample and the carrier is tight enough so that the variance of leakage currents through the aperture during electrical measurements lies below the signals to be measured by a factor of 5–

10.

29. A method for analyzing membranous samples such as cells, vesicles, and cellular organelles, comprising:

selecting a multiaperture system according to claim 1;

measuring at least one of an electrical potential difference and a current across at least a portion of the membranous sample sealed across one of the plurality of carriers, using the electrodes.

30. The method of claim 29, the membranous sample being a first membranous sample, further comprising:

sealing a second membranous sample across a second one of the fluid compartments; and

measuring at least one of an electrical potential difference and a current across at least a portion of the second sample, using the electrodes.

31. The method of claim 30, wherein the steps of measuring are performed sequentially for the first and second samples.

32. The method of claim 30, wherein the steps of measuring are performed simultaneously for the first and second samples.

33. A multiaperture system for analyzing membranous samples, comprising:

a holder,

a plurality of measurement sites disposed in the holder, each measurement site having a carrier that separates from one another two fluid compartments, the carrier containing an aperture that connects the two compartments, and a substantially planar seal surface circling the aperture; and

a membranous mammalian-cell-sized sample, selected from the group consisting of cells, organelles, and vesicles, sealed across an aperture at one of the measurement sites within the seal surface, wherein the seal between the sample and the carrier is tight enough so that the variance of leakage currents through the aperture during electrical measurements lies below the signals to be measured by a factor of 5–

10; and

wherein each compartment is in electrical contact with at least one electrode, the electrodes being adapted to apply and/or measure at least one of an electrical potential and a current, across at least a portion of the sample sealed across the aperture.

34. The multiaperture system of claim 33, wherein the seal has an electrical resistance of greater than about 1 GΩ

.

35. The system of claim 33, wherein the variance of leakage currents lies below the signals to be measured by a factor of 5–

10 when the signals to be measured lie between about 0.1 pA and about 50 pA at V_M=−

60 mV.

36. The system of claim 33, wherein the membranous sample is selected from the group consisting of cells, vesicles, and biological organelles.

37. The system of claim 33, wherein the system is configured for sequential measurements over at least two measurement sites.

38. The system of claim 33, wherein the system is configured for parallel measurements over at least two measurement sites.

39. The system of claim 33, further comprising a sample-handling mechanism for adding, manipulating, exchanging, storing, and/or removing samples and sample components from at least one compartment.

40. The system of claim 33, wherein each carrier is at least substantially planar.

41. The system of claim 33, wherein each carrier is formed at least in part of materials selected from the group consisting of silicon, silicon derivatives, glass, and plastic.

42. The system of claim 33, wherein the system includes a row of 8, 12, 16, 24, 32, or 48 measurement sites.

43. The system of claim 33, the holder being substantially rectangular, wherein the length of the holder ranges between about 125 mm and about 130 mm, and wherein the width of the holder ranges between about 80 mm and about 90 mm.

44. The system of claim 33, wherein the measurement sites correspond at least in part to microplate wells.

45. A method for analyzing membranous samples such as cells, vesicles, and cellular organelles, comprising:

selecting a multiaperture system according to claim 33;

measuring at least one of an electrical potential difference and a current across at least a portion of the membranous sample sealed across one of the plurality of carriers, using the electrodes.

46. The multiaperture system of claim 33, wherein the membranous mammalian-cell-sized sample is a mammalian cell.

47. The multiaperture system of claim 33, wherein each measurement site is substantially planar.

48. The multiaperture system of claim 47, wherein each measurement site is planar.

49. A multiaperture system for analyzing membranous samples selected from the group consisting of cells, vesicles, and cellular organelles, comprising:

a membranous mammalian-cell-sized sample, selected from the group consisting of cells, organelles, and vesicles;

a carrier having a plurality of apertures and substantially planar seal surfaces circling respective apertures, wherein the sample is sealed across at least one of the apertures and within the respective seal surface;

at least three fluid compartments, each fluid compartment connected to another of the at least three fluid compartments on an opposite side of the carrier via at least one of the apertures; and

at least three electrodes, each electrode in electrical contact with at least one of the fluid compartment, wherein the electrodes are adapted to apply and/or measure at least one of an electrical potential and a current, across at least a portion of the at least one of the samples, alter it is sealed across the at least one of the apertures;

wherein the fluid compartments and electrodes are separated on at least one side of the carrier to allow independent measurements;

wherein the maximum diameter of the aperture at the seal surface is between about 0.3 μ

m and about 7 μ

m.

50. The system of claim 49, wherein the seal has an electrical resistance of greater than about 1 GΩ

.

51. The system of claim 49, wherein the membranous samples are selected from the group consisting of cells, vesicles, and biological organelles.

52. The system of claim 49, wherein the membranous samples are selected from the group consisting of natural, artificial, and modified lipid membranes.

53. The system of claim 49, wherein at least one electrode is used for measurement over more than one aperture.

54. The system of claim 49, wherein several or all of the electrodes on one side of the carrier have a common electrical potential.

55. The system of claim 49, the carrier having first and second sides, wherein one compartment is disposed on the first side of the carrier, and the remaining compartments are disposed on the second side of the carrier.

56. The system of claim 49, wherein the system is configured for sequential measurements over at least two apertures.

57. The system of claim 49, wherein the system is configured for parallel measurements over at least two apertures.

58. The system of claim 49, further comprising a sample-handling mechanism for adding, manipulating, exchanging, storing, and/or removing samples and sample components from at least one compartment.

59. The system of claim 58, the carrier having first and second sides, wherein the sample-handling mechanism is configured to add and/or store fluid on one or both of the first and second sides of the carrier.

60. The system of claim 58, the carrier having first and second sides, wherein the sample-handling mechanism is configured to add cells, vesicles, and other biological organelles or particles on one or both of the first and second sides of the carrier.

61. The system of claim 58, wherein the sample-handling mechanism is configured to check the fluid level in the compartments.

62. The system of claim 58, wherein the sample-handling system is configured to introduce samples into a compartment through an inlet opening arranged concentrically above the aperture.

63. The system of claim 49, wherein the carrier is at least substantially planar.

64. The system of claim 49, wherein the carrier has an electrically charged surface that attracts biological membranes.

65. The system of claim 49, wherein the carrier has a surface that provides a molecule-specific bonding of cells, vesicles, or biological organelles on it.

66. The system of claim 49, wherein the carrier is formed at least in part of materials selected from the group consisting of silicon, silicon derivatives, glass, and plastic.

67. The system of claim 49, wherein the carrier comprises a silicon chip or a silicon wafer.

68. The system of claim 49, the carrier having first and second sides, wherein the compartments on at least one of the first and second side of the carrier are open.

69. The system of claim 49, wherein at least two fluid compartments are disposed on one side of the carrier, each of those compartments being in contact, via the apertures, with a single fluid compartment disposed on the other side of the carrier.

70. The system of claim 49, further comprising an optical-analysis mechanism configured for optical analysis samples positioned about at least one aperture.

71. The system of claim 70, wherein the optical-analysis mechanism is configured for fluorescence measurements.

72. The system of claim 49, further comprising a holder adapted to support the carrier.

73. The system of claim 49, wherein the system includes a row of 8, 12, 16, 24, 32, or 48 apertures.

74. The system of claim 49, wherein the application of a voltage difference, mediated by the electrodes, creates an inhomogeneous electric field around the aperture, which becomes larger as the aperture is approached.

75. A method for analyzing samples such as cells, vesicles, and cellular organelles, comprising:

selecting a multiaperture system according to claim 49;

adding a sample to one of the fluid compartments; and

measuring at least one of an electrical potential difference and a current across at least a portion of the sample, using the electrodes.

76. The method of claim 75, further comprising:

adding a second sample to a second one of the fluid compartments; and

measuring at least one of an electrical potential difference and a current across at least a portion of the second sample, using the electrodes.

77. The method of claim 76, the sample being a first sample, where the steps of measuring are performed sequentially for the first and second samples.

78. The method of claim 76, the sample being a first sample, where the steps of measuring are performed simultaneously for the first and second samples.

79. The A multiaperture system of claim 49, wherein the carrier is substantially planar.

80. The A multiaperture system of claim 79, wherein the carrier is planar.

81. A multiaperture system for analyzing membranous samples selected from the group consisting of cells, vesicles, and cellular organelles, comprising:

a membranous mammalian-cell-sized sample, selected from the group consisting of cells, organelles, and vesicles;

a holder; and

a plurality of measurement sites disposed in the holder, each measurement site having a carrier that separates from one another two fluid compartments;

wherein the carrier contains an aperture that connects the two compartments and a substantially planar seal surface circling the aperture, wherein the sample is sealed across the aperture and within the seal surface;

wherein each compartment is in electrical contact with at least one electrode, the electrodes being adapted to measure at least one of an electrical potential and a current, across at least a portion of the sample, after the sample has been sealed across the aperture; and

wherein the fluid compartments and electrodes are separated on at least one side of the carrier to allow independent measurements;

wherein the maximum diameter of the aperture at the seal surface is between about 0.3 μ

m and about 7 μ

m.

82. The multiaperture system of claim 81, wherein the seal has an electrical resistance of greater than about 1 Ω

.

83. The system of claim 81, wherein each fluid compartment is in electrical contact with at least one electrode.

84. The multiaperture system of claim 81, wherein the measurement sites each include only a single aperture.

85. The system of claim 81, wherein the membranous samples are selected from the group consisting of cells, vesicles, and biological organelles.

86. The system of claim 81, the carrier having first and second sides, wherein the fluid compartments and electrodes are separated on the first side of the carrier, and wherein there is a single fluid compartment on the second side of the carrier.

87. The system of claim 81, wherein the system is configured for sequential measurements over at least two measurement sites.

88. The system of claim 81, wherein the system is configured for parallel measurements over at least two measurement sites.

89. The system of claim 81, further comprising a sample-handling mechanism for adding, manipulating, exchanging, storing, and/or removing samples and sample components from at least one compartment.

90. The system of claim 81, wherein each carrier is at least substantially planar.

91. The system of claim 81, wherein each carrier is formed at least in part of materials selected from the group consisting of silicon, silicon derivatives, glass, and plastic.

92. The system of claim 81, wherein the system includes a row of 8, 12, 16, 24, 32, or 48 measurement sites.

93. The system of claim 81, the holder being substantially rectangular, wherein the length of the holder ranges between about 125 mm and about 130 mm, and wherein the width of the holder ranges between about 80 mm and about 90 mm.

94. The system of claim 81, wherein the measurement sites correspond at least in part to microplate wells.

95. A method for analyzing membranous samples such as cells, vesicles, and cellular organelles, comprising:

selecting a multiaperture system according to claim 81;

adding a sample to one of the fluid compartments; and

measuring at least one of an electrical potential difference and a current across at least a portion of the membranous sample sealed across one of the plurality of carriers, using the electrodes.

96. The A multiaperture system of claim 81, wherein the carrier is substantially planar.

97. The A multiaperture system of claim 96, wherein the carrier is planar.