Microlithographic array for macromolecule and cell fractionation

• US 5,837,115 A
• Filed: 06/07/1995
• Issued: 11/17/1998
• Est. Priority Date: 06/08/1993
• Status: Expired due to Term

• Current Assignee: BTG International Inc. (Boston Scientific Corporation)
• Original Assignee: British Technology Group Ltd., Cornell Research Foundation Incorporated (Cornell University)
• Inventors: Volkmuth, Wayne D., Rathbun, Lynn C., Austin, Robert H.
• Primary Examiner(s): Gorgos, Kathryn L.
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/474,688
• Time in Patent Office: 1,259 Days
• Field of Search: 435/283.1, 209/156, 209/155, 204/600, 204/450
• US Class Current: 204/450
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: G01N 15/0255 with mechanical, e.g. inertial, classification, and investigation of sorted collections with centrifuges G01N15/042
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 30/6095 Micromachined or nanomachined, e.g. micro- or nanosize
• CPC Class Codes: G01N 30/92 Construction of the plate

Abstract

A sorting apparatus and method for fractionating and simultaneously viewing individual microstructures, such as free cells, viruses, macromolecules, or minute particles in a fluid medium. The sorting apparatus is composed of a substrate having a receptacle located therein, the receptacle having sidewalls and a floor. An array of obstacles is positioned within the receptacle with the obstacles upstanding from the floor of the receptacle. A transparent cover overlies the array of obstacles to cover the receptacle and afford visual observation of migration of the microstructures exclusively through the array of obstacles. Electrodes may be positioned within the receptacle to generate an electric field in the fluid medium in the receptacle in order to induce the migration of the microstructures. Migration of the microstructures may also occur, for example, by a hydrodynamic field, an optical field, a magnetic field, or a gravity field applied to the receptacle. The obstacles of the array of obstacles may be of various shapes such as round posts, rectangular bunkers, or v-shaped or cup-shaped structures. The arrays of obstacles are formed of a predetermined and reproducible pattern, and can be reused. Methods for manufacturing and using the apparatus are also claimed.

Claims

1. An apparatus for sorting microstructures, in a fluid medium, said apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;

b. a two-dimensional array of obstacles positioned in said receptacle intermediate said first and second ends thereof; and

c. ceiling means positioned over and contacting said array of obstacles for precluding migration of the microstructures between said array of obstacles and said ceiling means.

2. An apparatus as recited in claim 1, wherein said obstacles are upstanding from said floor of said receptacle in a plurality of rows traversing said migration direction.

3. A sorting apparatus as recited in claim 2, wherein each of said obstacles has a top at the end thereof opposite from said floor of said housing, and said top of each of said obstacles is permanently secured to said ceiling means.

4. A sorting apparatus as recited in claim 2, wherein said obstacles of said array are arranged in a preselected and reproducible pattern, each obstacle in said array of obstacles having a height measured in a direction normal to said floor of said receptacle, a length measured in a direction parallel to said migration direction, and a width measured in a direction normal to said migration direction, and wherein the distance of each of said obstacles from an adjacent of said obstacles defines a separation distance therebetween.

5. A sorting apparatus as recited in claim 4, wherein said height of each of said obstacles is in a range of from about 0.01 microns to about 20.0 microns.

6. A sorting apparatus as recited in claim 4, wherein said height of each of said obstacles is in a range of from about 0.01 microns to about 0.50 microns.

7. A sorting apparatus as recited in claim 4, wherein said height of each of said obstacles is in a range of from about 1.0 micron to about 5.0 microns.

8. A sorting apparatus as recited in claim 4, wherein said separation distance is substantially equal to a radius of gyration of each of the microstructures.

9. A sorting apparatus as recited in claim 4, wherein said separation distance is in a range of from about 0.01 microns to about 50.0 microns.

10. A sorting apparatus as recited in claim 4, wherein said separation distance is in a range of from about 0.01 microns to about 1.0 micron.

11. A sorting apparatus as recited in claim 4, wherein said separation distance is in a range of from about 1.0 micron to about 20.0 microns.

12. A sorting apparatus as recited in claim 4, wherein said separation distance is substantially equal to said length of said obstacles.

13. A sorting apparatus as recited in claim 4, wherein said obstacles comprise posts.

14. A sorting apparatus as defined in claim 4, wherein said obstacles comprise elongated bunkers.

15. A sorting apparatus as recited in claim 14, wherein the longitudinal axis of said bunkers are disposed in alignment with said migration direction of said receptacle.

16. A sorting apparatus as defined in claim 4, wherein said obstacles comprise elongated interconnected bunkers forming an open lattice of nonlinear channels between said first and said second end of said receptacle.

17. A sorting apparatus as recited in claim 2, wherein the top of each of said obstacles is ion fusion bonded to said ceiling means.

18. An apparatus as recited in claim 2, wherein said obstacles in adjacent of said rows thereof are aligned along said migration direction.

19. An apparatus as recited in claim 2, wherein said obstacles in adjacent of said rows thereof are offset relative to said migration direction.

20. An apparatus as recited in claim 2, wherein said rows of said obstacles are parallel to each other.

21. An apparatus as recited in claim 2, wherein said rows of said obstacles are perpendicular to said migration direction.

22. An apparatus as recited in claim 2, wherein said rows of said obstacles are oriented at an angle to said migration direction.

23. An apparatus as recited in claim 22, wherein said angle has a measure of about 45°

.

24. An apparatus as recited in claim 2, wherein spaces between adjacent of said obstacles in a cross section of said array of said obstacles taken normal to said floor of said receptacle define pores of a lattice structure, and said pores of said lattice structure are uniform in size and shape throughout said array of said obstacles.

25. An apparatus as recited in claim 2, wherein spaces between adjacent of said obstacles in a cross section of said array make a normal to said floor of said receptacle define pores of a lattice structure, and said pores of said lattice structure vary in size and shape throughout said array.

26. A sorting apparatus as defined in claim 1, wherein said obstacles cumulatively comprise a percolating array.

27. A sorting apparatus as recited in claim 1, wherein said ceiling means comprises a coverslip extending across said receptacle from one of said pair of upstanding opposing side walls to the other.

28. A sorting apparatus as recited in claim 27, wherein said coverslip is transparent.

29. A sorting apparatus as recited in claim 1, wherein the height of said side walls of said receptacle as measured normal to said floor of said receptacle define the depth of said receptacle, and said receptacle is so configured that said depth thereof causes the microstructures to migrate through said receptacle in said migration direction in essentially a single layer.

30. An apparatus as recited in claim 1, wherein migration of the microstructures through said obstacles is in said migration direction.

31. An apparatus as recited in claim 1, wherein said obstacles are upstanding from said floor of said receptacle in a uniform pattern.

32. An apparatus as recited in claim 1, wherein said obstacles are upstanding from said floor of said receptacle in a staggered pattern.

33. A sorting apparatus for fractionating microstructures, in a fluid medium, said sorting apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;

b. sifting means positioned within said receptacle intermediate said first and second ends thereof traversing said migration direction for interacting with the microstructures to partially hinder migration of the microstructures in said migration direction in the fluid medium, said sifting means comprising an array of obstacles upstanding from said floor of said receptacle, and arranged in a preselected and reproducible pattern, each of said obstacles having;

(i) a v-shaped cross section in a plane disposed parallel to said floor of said receptacle, said v-shaped cross section of said obstacles comprising first and second arms intersecting at an end of each to form a vertex of said v-shaped cross section, the other ends of said first and second arms remote from said vertex forming an open end of said v-shaped cross section;

(ii) a top at the end thereof opposite from said floor of said receptacle;

(iii) a height measured in a direction normal to said floor of said receptacle;

(iv) a length measured in a direction parallel to said migration direction; and

(v) a width measured in a direction normal to said migration direction,

(vi) the distance of each of said obstacles from an adjacent of said obstacles defining a separation distance therebetween; and

c. ceiling means positioned over said sifting means for covering said receptacle and for causing migration of the microstructures in said receptacle to occur through said sifting means exclusively.

34. A sorting apparatus as recited in claim 33, wherein the open end of the v-shaped cross section of said obstacles is disposed opposing said migration direction of said receptacle.

35. A sorting apparatus for fractionating microstructures in a fluid medium, said sorting apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;

b. sifting means positioned within said receptacle intermediate said first and second ends thereof traversing said migration direction for interacting with the microstructures to partially hinder migration of the microstructures in said migration direction in the fluid medium, said sifting means comprising an array of obstacles upstanding from said floor of said receptacle and arranged in a preselected and reproducible pattern, each of said obstacles having;

(i) a cup-shaped cross section in a plane disposed parallel to said floor of said receptacle, said cup-shaped cross section of said obstacles comprising;

(A) a first leg and a second leg positioned in opposed relationship substantially parallel to said migration direction, each of said first and second legs having thereby oppositely disposed first and second ends; and

(B) a third leg positioned substantially perpendicular to said migration direction and being connected between said first ends of each of said first and second legs, thereby to define between said second ends of said first and second legs an open end of said cup-shaped cross section;

(ii) a top at the end thereof opposite from said floor of said receptacle;

(iii) a height measured in a direction normal to said floor of said receptacle;

(iv) a length measured in a direction parallel to said migration direction; and

(v) a width measured in a direction normal to said migration direction,

(vi) the distance of each of said obstacles from an adjacent of said obstacles defining a separation distance therebetween; and

c. ceiling means positioned over said sifting means for covering said receptacle and for causing migration of the microstructures in said receptacle to occur through said sifting means exclusively.

36. A sorting apparatus as recited in claim 35, wherein said open end of said cup-shaped cross section of said obstacles is disposed opposing said migration direction of said receptacle.

37. An apparatus for sorting microstructures in a fluid medium, said apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, the height of said side walls defining a depth of said receptacle;

b. a two-dimensional array of obstacles upstanding from said floor of said receptacle, each of said obstacles in said array having a height measured in a direction normal to said floor of said receptacle, said height of said obstacles being substantially equal to said depth of said receptacle; and

c. ceiling means positioned over said array of obstacles for covering said receptacle and for causing the migration of the microstructures within said receptacle to occur exclusively through said array of obstacles, the end of each of said obstacles remote from said floor of said receptacle engaging said ceiling means, thereby to preclude migration of the microstructures between said ceiling means and each of said obstacles.

38. A sorting apparatus as recited in claim 37, wherein said ceiling means comprises a coverslip extending across said receptacle from one of said pair of upstanding opposing side walls to the other.

39. A sorting apparatus as recited in claim 38, wherein said coverslip and said substrate are comprised of materials having substantially similar coefficients of thermal expansion.

40. An apparatus as recited in claim 38, wherein said coverslip is comprised of a ceramic.

41. An apparatus as recited in claim 38, wherein said coverslip is comprised of quartz.

42. An apparatus as recited in claim 38, wherein said coverslip is comprised of sapphire.

43. An apparatus as recited in claim 38, wherein said coverslip is comprised of pyrex.

44. An apparatus as recited in claim 38, wherein said coverslip is comprised of silicon.

45. A sorting apparatus as recited in claim 38, wherein said coverslip is transparent.

46. A sorting apparatus as recited in claim 38, wherein said coverslip is ion fusion bonded to said pair of upstanding opposing side walls.

47. An apparatus as recited in claim 37, wherein said substrate is comprised of a material that can be subjected to photolithographic etching.

48. An apparatus as recited in claim 37, wherein said substrate is comprised of silicon.

49. An apparatus as recited in claim 37, wherein said substrate is comprised of quartz.

50. An apparatus as recited in claim 37, wherein said substrate is comprised of sapphire.

51. A sorting apparatus as recited in claim 37, wherein said depth of said receptacle and said height of said obstacles is such that when the microstructures are caused to migrate in the fluid medium from said first end of said receptacle to said second end of said receptacle, the microstructures do so in essentially a single layer.

52. A sorting apparatus as recited in claim 37, wherein said height of each of said obstacles is in a range of from about 0.01 microns to about 20.0 microns.

53. A sorting apparatus as recited in claim 37, wherein said height of said obstacles is in a range of from about 0.01 microns to about 0.50 microns.

54. A sorting apparatus as recited in claim 37, wherein said height of said obstacles is in a range of from about 1.0 micron to about 5.0 microns.

55. A lattice structure of predetermined pore size and shape for use in sorting individual microstructures in a fluid medium, said lattice structure comprising:

a. a substrate having thereon a planar surface, said planar surface defining a floor of said lattice structure;

b. a coverslip disposed parallel to and separated from said floor, thereby to define between said coverslip and said floor a migration channel capable of receiving the microstructures in the fluid medium for migration of the microstructures through said migration channel;

c. a two-dimensional array of obstacles extending between and contacting said floor and said coverslip in a preselected and reproducible pattern, the space between adjacent of said obstacles in a cross section of said array taken normal to said floor of said lattice structure defining pores of said lattice structure, said pores of said lattice structure being so sized and configured as to slow the rate of migration of the microstructures.

56. A lattice structure as recited in claim 55, wherein said obstacles are integrally formed with said substrate.

57. A lattice structure as recited in claim 55, wherein said substrate, said coverslip, and said obstacles are comprised of materials having substantially similar coefficients of thermal expansion.

58. A lattice structure as recited in claim 56, wherein said obstacles are integrally formed at one end thereof with said substrate and bonded at the other end thereof to said coverslip.

59. A lattice structure as recited in claim 55, wherein said pores of said lattice structure assume a generally rectangular shape.

60. A lattice structure as recited in claim 55, wherein said coverslip is transparent.

61. A lattice structure as recited in claim 55, wherein said coverslip affords visual observation of the migration of the microstructures.

62. A lattice structure as recited in claim 55, wherein each of said obstacles in said array has a top at the end thereof opposite from said floor of said lattice structure, and said coverslip is ion fusion bonded to said top of each of said obstacles in said array.

63. A lattice structure as recited in claim 55, wherein the depth of said migration channel is defined by the distance between said floor and said cover slip measured normal to said floor, and said depth of said migration channel is such as to cause migration of the microstructures in said migration channel to occur in essentially a single layer.

64. A lattice structure as recited in claim 55, wherein said pores of said lattice structure are so configured as to cause the microstructures to migrate through said lattice structure in essentially a single file.

65. A lattice structure as recited in claim 64, wherein said pores of said lattice structure are so configured as to cause the microstructures to migrate through said lattice structure in essentially a single file.

66. A lattice structure as recited in claim 55, wherein each obstacle in said array of obstacles has a height measured in a direction normal to said floor of said migration channel, and said height of each of said obstacles is in a range of from about 0.01 microns to about 20.0 microns.

67. A lattice structure as recited in claim 55, wherein each obstacle in said array of obstacles has a height measured in a direction normal to said floor of said migration channel, and said height of each of said obstacles is in a range of from about 0.01 microns to about 0.50 microns.

68. A lattice structure as recited in claim 55, wherein each obstacle in said array of obstacles has a height measured in a direction normal to said floor of said migration channel, and said height of each of said obstacles is in a range of from about 1.0 micron to about 5.0 microns.

69. A lattice structure as recited in claim 55, wherein said array of obstacles comprise a plurality of rows of said obstacles traversing said migration channel.

70. A lattice structure as recited in claim 69, wherein said rows of obstacles are parallel to each other.

71. A lattice structure as recited in claim 55, wherein said obstacles are disposed in a uniform pattern.

72. A lattice structure as recited in claim 55, wherein said obstacles is disposed in a staggered pattern.

73. A lattice structure as recited in claim 55, wherein said pores of said lattice structure are uniform in size and shape throughout said array of said obstacles.

74. A lattice structure as recited in claim 55, wherein said migration channel has a depth measured between said floor of said lattice structure and said coverslip in a range from about 0.01 microns to less than about 0.1 microns.

75. A lattice structure as recited in claim 55, wherein said migration channel has a depth measured between said floor of said lattice structure and said coverslip in a range from about 0.01 microns to about 0.06 microns.

76. A reusable apparatus for sorting individual microstructures in a fluid medium, said apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the microstructures in the fluid medium, whereby when the microstructures are caused to migrate in the fluid medium from said first end through said second end of said receptacle the microstructures do so in essentially a single layer; and

b. a two-dimensional array of obstacles positioned in said receptacle intermediate said first and second ends thereof, said obstacles being comprised of a rigid self-sustaining material that is chemically inert to the microstructures and to the fluid medium in a normal range of temperatures suitable for fractionating the microstructures; and

c. capping means positioned over and contacting said array of obstacles for affording visual observation of migration of the microstructures in said migration direction through said receptacle, and for precluding migration of the microstructures between said array of obstacles and said capping means.

77. A sorting apparatus as recited in claim 76, wherein said substrate and said array of obstacles are integrally formed.

78. A sorting apparatus as recited in claim 76, wherein said substrate and said array of obstacles are comprised of a material that can be subjected to photolithographic etching.

79. A sorting apparatus as recited in claim 76, wherein said substrate and said array of obstacles are comprised of silicon.

80. A sorting apparatus as recited in claim 76, wherein each of said obstacles has a height measured normal to said floor of said receptacle, and said height is substantially equal to said depth of said receptacle.

81. A sorting apparatus as recited in claim 76, wherein migration of the microstructures through said hindered environment means is in said migration direction.

82. An apparatus for channelling cells through passageways and simultaneously viewing the cells in a fluid medium, said apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having a floor bound by a pair of upstanding opposing side walls and a first end and a second end, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the cells in the fluid medium, whereby when the cells are caused to migrate in the fluid medium through said receptacle, the cells do so in essentially a single layer, the migration of cells from the first end to the second end of said receptacle defining a migration direction for said receptacle;

b. a two-dimensional array of obstacles positioned in said receptacle intermediate said first and second ends thereof, said array of obstacles allowing passage of the cells through said receptacle in at least one single file; and

c. capping means for covering said receptacle intermediate said first and second ends thereof and for affording visual observation of the migration of the cells through said array of obstacles and through said receptacle.

83. An apparatus as recited in claim 82, wherein passageways are defined between adjacent ones of said obstacles.

84. An apparatus as recited in claim 83, wherein said passageways are linear.

85. An apparatus as recited in claim 83, wherein said passageways are in alignment with said migration direction.

86. An apparatus as recited in claim 83, wherein each of said passageways has a width in the range of from about 1.0 micron to about 10.0 microns.

87. An apparatus as recited in claim 83, wherein each of said passageways has a depth in the range of from about 1.0 micron to about 10.0 microns.

88. An apparatus for channelling cells through passageways and simultaneously viewing the cells in a fluid medium, said apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having a floor bound by a pair of upstanding opposing side walls and a first end and a second end, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the cells in the fluid medium, whereby when the cells are caused to migrate in the fluid medium through said receptacle, the cells do so in essentially a single layer, the migration of cells from the first end to the second end of said receptacle defining a migration direction for said receptacle;

b. channeling means for allowing passage of cells through said receptacle in at least one single file;

c. capping means for covering said receptacle intermediate said first and second ends thereof and for affording visual observation of the migration of the cells through said channeling means and through said receptacle; and

d. means for measuring the amount of energy consumed during passage of the cells through said channeling means.

89. An apparatus for channelling cells through passageways and simultaneously viewing the cells in a fluid medium, said apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having a floor bound by a pair of upstanding opposing side walls and a first end and a second end, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the cells in the fluid medium, whereby when the cells are caused to migrate in the fluid medium through said receptacle, the cells do so in essentially a single layer, the migration of cells from the first end to the second end of said receptacle defining a migration direction for said receptacle;

b. channeling means for allowing passage of cells through said receptacle in at least one single file, said channeling means comprising passageways that are curved and are positioned within said receptacle; and

c. capping means for covering said receptacle intermediate said first and second ends thereof and for affording visual observation of the migration of the cells through said channeling means and through said receptacle.

90. A method for manufacturing a sorting apparatus for fractionating and simultaneously viewing individual microstructures in a fluid medium, said method comprising the steps of:

a. forming on one side of a substrate a receptacle having a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle, said depth of said receptacle being commensurate with the size of the microstructures in the fluid medium, whereby when the microstructures are caused to migrate in the fluid medium through said receptacle the microstructures do so in essentially a single layer;

b. building a two-dimensional array of upstanding obstacles in a preselected and reproducible pattern on said floor of said receptacle, the end of each of said obstacles remote from said floor defining the top of said obstacle; and

c. engaging a transparent coverslip to said tops of said obstacles, thereby to preclude migration of the microstructures between said coverslip and said tops of obstacles.

91. A method as recited in claim 90, wherein said steps of forming and building are performed simultaneously.

92. A method as recited in claim 90, wherein said step of building comprises the steps of:

a. positioning a photoresist layer on the surface of said substrate over areas of said substrate corresponding to said tops of said obstacles of said arrays;

b. etching said substrate to a preselected depth equal to said depth of said receptacle; and

c. dissolving said photoresist layer from said substrate.

93. A method as defined in claim 92, wherein said step of etching comprises the step of ion milling said surface of said substrate with said photoresist layer thereon.

94. A method as defined in claim 92, wherein said step of etching comprises the step of chemical etching said surface of said substrate with said photoresist layer thereon.

95. A method as recited in claim 90, wherein said step of securing comprises the steps of:

a. positioning said coverslip over said array of obstacles such that said coverslip is in contact with said tops of said obstacles; and

b. applying an electric field between said coverslip and said array of obstacles, thereby causing ions to migrate therebetween.

96. A method as recited in claim 95, wherein the applied voltage in said step of applying is in a range of from 200 V to 2000 V.

97. A method as recited in claim 95, wherein the applied voltage is said step of applying is 1 kilovolt.

98. A method as recited in claim 95, wherein the step of applying is conducted in a temperature in a range of from 300°

C. to 600°

C.

99. A method for sorting microstructures comprising the steps of:

a. placing the microstructures into a fluid medium;

b. introducing the fluid medium with the microstructures therein into a receptacle having a first end and a second end and a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle;

c. positioning within said receptacle a two-dimensional array of obstacles disposed in a preselected and reproducible pattern; and

d. inducing the microstructures in the fluid to migrate through said array of obstacles positioned within said receptacle.

100. A method as recited in claim 99, wherein said step of inducing comprises the step of imposing a hydrodynamic field on said fluid medium.

101. A method as recited in claim 99, wherein said step of inducing comprises the step of imposing a gravity field on the microstructures.

102. A method as recited in claim 99, further comprising the step of visually observing migration of the microstructures through said array of obstacles.

103. A method as recited in claim 99, wherein said depth of said receptacle is such that when the microstructures migrate in the fluid medium through said receptacle, the microstructures do so in essentially a single layer.

104. A method as recited in claim 99, wherein said step of inducing comprises the step of imposing an electric field through said fluid medium on the microstructures.

105. A method for sorting microstructures, said method comprising the steps of;

a. placing the microstructures into a fluid medium;

b. introducing the fluid medium with the microstructures therein into a receptacle having a first end and a second end and a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle;

c. positioning within said receptacle an array of upstanding obstacles in a preselected and reproducible pattern; and

d. inducing the microstructures in the fluid to migrate through said array of obstacles positioned within said receptacle, said step of inducing comprising the steps of;

(i) positioning a first electrode on said floor of said receptacle at said first end of said receptacle;

(ii) positioning a second electrode on said floor of said receptacle at said second end of said receptacle; and

(iii) coupling a power source between said first and second electrodes.

106. A method as recited in claim 105, further comprising the steps of:

a. positioning a pair of sensing electrodes at first and second preselected points in said array of obstacles;

b. amplifying the signal obtained on each of said sensing electrodes; and

c. differentiating the signals obtained in said step of amplifying to produce an output signal corresponding to the intensity of said electric field in said array between said first and second predetermined points.

107. A method for sorting microstructures, said method comprising the steps of:

a. placing the microstructures into a fluid medium;

b. introducing the fluid medium with the microstructures therein into a receptacle having a first end and a second end and a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle;

c. positioning within said receptacle an array of upstanding obstacles in a preselected and reproducible pattern; and

d. inducing the microstructures in the fluid to migrate through said array of obstacles positioned within said receptacle, said step of inducing comprising the step of focusing light onto the microstructures.

108. A method for sorting microstructures, said method comprising the steps of:

a. placing the microstructures into a fluid medium;

b. introducing the fluid medium with the microstructures therein into a receptacle having a first end and a second end and a floor bound by a pair of upstanding opposing side walls, the height of said side walls defining a depth of said receptacle;

c. positioning within said receptacle an array of upstanding obstacles in a preselected and reproducible pattern; and

d. inducing the microstructures in the fluid to migrate through said array of obstacles positioned within said receptacle, said step of inducing comprising the step of imposing a magnetic field on the microstructures.

109. A sorting apparatus for fractionating microstructures in a fluid medium, said sorting apparatus comprising:

a. a substrate with a receptacle formed in a side thereof, said receptacle having a first end, a second end, and a floor bound on opposite sides by a pair of side walls that extend between said first end and second end of said receptacle, migration of the microstructures from said first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle;

b. a two-dimensional array of obstacles upstanding from said floor of said receptacle in a preselected and reproducible pattern, the spaces between adjacent of said obstacles in a cross section of said array taken normal to said floor of said receptacle defining pores of a lattice structure capable of interacting with the microstructures to differentially slow the rate of migration of individual types of the microstructures during migration of the microstructures in said migration direction through said array of obstacles, each of said pores of said lattice structure having dimensions comprising;

i. a height of said pores measured in a direction normal to said floor of said receptacle, said height of said pores of said lattice structure being in a range of from about 0.01 microns to about 20.0 microns; and

ii. a separation distance of said pores measured parallel to said floor of said receptacle between adjacent of said obstacles, said separation distance of said pores of said lattice structure being in a range of from about 0.01 microns to about 50.0 microns; and

c. a coverslip extending across said receptacle from one of said side walls to the other and contacting the end of each of said obstacles opposite from said floor of said receptacle.

110. A sorting apparatus as recited in claim 109, wherein said separation distance of said pores of said lattice structure is in a range of from about 0.01 microns to about 1.0 micron.

111. A sorting apparatus as recited in claim 109, wherein said separation distance of said pores of said lattice structure is in a range of from about 1.0 micron to about 20.0 microns.

112. A sorting apparatus as recited in claim 109, wherein said separation distance of said pores of said lattice structure is in a range of from about 1.0 micron to about 10.0 microns.

113. A sorting apparatus as recited in claim 109, wherein said height of said pores of said lattice structure is in a range of from about 1.0 micron to about 10.0 microns.

114. A sorting apparatus as recited in claim 109, wherein said height of said pores of said lattice structure is in a range of from about 0.01 micron to about 0.50 microns.

115. A sorting apparatus as recited in claim 109, wherein said height of said pores of said lattice structure is in a range of from about 1.0 micron to about 5.0 microns.

116. A sorting apparatus as recited in claim 109, wherein said height of said pores of said lattice structure is in a range from about 0.01 microns to less than about 0.1 microns.

117. A sorting apparatus as recited in claim 109, wherein said height of said pores of said lattice structure is in a range from about 0.01 microns to about 0.06 microns.

118. A sorting apparatus for fractionating microstructures in a fluid medium, said sorting apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle; and

b. sifting means positioned within said receptacle intermediate said first and second ends thereof traversing said migration direction for interacting with the microstructures to partially hinder migration of the microstructures in said migration direction in the fluid medium, said sifting means comprising an array of obstacles upstanding from said floor of said receptacle and arranged in a preselected and reproducible pattern, each of said obstacles having a v-shaped cross section in a plane disposed parallel to said floor of said receptacle, said v-shaped cross section of said obstacles comprising;

(i) a first arm and

(ii) a second arm disposed at an angel to said first arm and intersecting said first arm at the vertex of said v-shaped cross section, the ends of said first and second arms remote from said vertex forming the open end of said v-shaped cross section.

119. A sorting apparatus as recited in claim 118, wherein said open end of said v-shaped cross section of said obstacles is disposed opposing said migration direction of said receptacle.

120. A sorting apparatus for fractionating microstructures in a fluid medium, said sorting apparatus comprising:

a. a substrate having a receptacle located on a side thereof, said receptacle having first and second ends and a floor bound on opposite sides by a pair of upstanding opposed side walls extending between said first and second ends of said receptacle, migration of the microstructures from the first end of said receptacle to said second end of said receptacle defining a migration direction for said receptacle; and

b. sifting means positioned within said receptacle intermediate said first and second ends thereof traversing said migration direction for interacting with the microstructures to partially hinder migration of the microstructures in said migration direction in the fluid medium, said sifting means comprising an array of obstacles upstanding from said floor of said receptacle and arranged in a preselected and reproducible pattern, each of said obstacles having a cup-shaped cross section in a plane disposed parallel to said floor of said receptacle, said cup-shaped cross section of said obstacles comprising;

(i) a first leg and a second leg positioned in opposed relationship substantially parallel to said migration direction, each of said first and second legs having thereby oppositely disposed first and second ends; and

(ii) a third leg positioned substantially perpendicular to said migration direction and being connected between said first ends of each of said first and second legs, thereby to define between said second ends of said first and second legs the open end of said cup-shaped cross section.

121. A sorting apparatus as recited in claim 120, wherein said open end of said cup-shaped cross section of said obstacles is disposed opposing said migration direction of said receptacle.