Claims
- 1. A device for preparing a reaction substrate for conducting multiplexed microassays to determine bindings between a target analyte and a capture probe, including:
a plurality of capillary tubes, each tube having a proximal end and a distal end; an attachment site for holdings said capillary tubes at a point spaced from the distal ends of such capillary tubes; an array template for slidably holding each capillary tube near its distal end, and for allowing the distal end of each capillary tube to move with respect to the attachment site; at least one manifold for positioning the proximal end of each capillary tube within a corresponding supply chamber, wherein each supply chamber is capable of supplying a liquid reagent to at least one corresponding capillary tube; and a positioning device for precisely positioning the array template and said capillary tubes with respect to said reaction substrate and depositing liquid reagents from said capillary tubes onto said reaction substrate as biosites.
- 2. The device of claim 1, wherein said plurality of capillary tubes comprises about 2 to about 10,000 tubes, having a center to center spacing of about 80 μm to about 5 mm.
- 3. The device of claim 1, wherein each supply chamber supplies only one capillary tube.
- 4. The device of claim 1, wherein a supply chamber supplies a plurality of capillary tubes.
- 5. The device of claim 1, wherein said capillary tubes comprise stainless steel, plastic, rubber, glass, or fused silica coated with polyimide.
- 6. The device of claim 1, wherein said device includes a plurality of array templates.
- 7. The device of claim 1, further comprising a positioning device for precisely positioning said reaction substrate.
- 8. The device of claim 1, wherein said capillaries have an inside diameter of about 10 to about 200 μm, and an outside diameter of about 80 to about 500 μm.
- 9. The device of claim 1, wherein said array template comprises an array of sleeves, each sleeve having an inside diameter sufficient to permit a capillary to slide through, and a length sufficient to allow a precise pattern to be maintained while depositing fluids onto said reaction substrate.
- 10. The device of claim 1, wherein said array template comprises a plurality of holes formed in a rigid material.
- 11. The device of claim 1, wherein said array template comprises a rigidly formed or held mesh.
- 12. The device of claim 1, further comprising a housing for containing said supply chambers.
- 13. The device of claim 12, wherein said housing is capable of maintaining an inert atmosphere.
- 14. The device of claim 1, wherein said housing is capable of maintaining an elevated or reduced temperature.
- 15. The device of claim 12, wherein said housing may be pressurized to a predetermined pressure.
- 16. The device of claim 15, wherein said pressure is modulated to control the flow of liquid reagents through said capillary tubes.
- 17. The device of claim 1, wherein said supply chambers are positioned higher than the distal end of said capillary tubes to provide a pressure head.
- 18. The device of claim 1, wherein deposit of capture probe is controlled by electrophoresis.
- 19. The device of claim 1, wherein supply of liquid reagent is controlled by electro-osmosis.
- 20. The device of claim 1, wherein said device includes multiple sets of supply chambers, each for supplying liquid reagent to a subset of said capillary tubes.
- 21. The device of claim 1, wherein said capillary tubes are free to flex between said attachment site and said array template.
- 22. The device of claim 1, wherein the reaction substrate is pre-etched to define a pattern of reactive areas matching the geometry of the deposited biosites.
- 23. The device of claim 1, wherein said biosites are deposited substantially simultaneously.
- 24. The device of claim 1, wherein each biosite is fluidically isolated from each other biosite.
- 25. A reaction substrate for conducting multiplexed microassays to determine binding of a target molecule and a capture probe/target probe complex, said reaction substrate including an array of biosites, each biosite comprising a single type of capture probe bound to said substrate, each capture probe capable of binding to a corresponding target probe having a capture probe specific domain which specifically binds with a corresponding capture probe, and a target analyze specific domain which specifically binds with a target analyte.
- 26. The reaction substrate of claim 25, wherein said reaction substrate is about 50 μm to about 300 μm in thickness.
- 27. The reaction substrate of claim 25, wherein each biosite on the reaction substrate comprises a capture probe different from the capture probe in every other biosite on said reaction substrate.
- 28. The reaction substrate of claim 25, wherein said array of biosites comprises from about 2 to about 10,000 biosites.
- 29. The reaction substrate of claim 25, further including a target probe having a capture probe specific domain which specifically binds with a corresponding capture probe, and a target analyte specific domain which specifically binds with a target analyte.
- 30. The reaction substrate or claim 29, wherein each capture probe is a first oligonucleotide, each capture probe specific domain is a second oligonucleotide, and each target analyte specific domain is a third oligonucleotide.
- 31. The reaction substrate of claim 29, wherein each capture probe is a hapten, each capture probe specific domain is a hapten binding polypeptide, and each target analyte specific domain is an oligonucleotide.
- 32. The reaction substrate of claim 31, wherein each hapten binding polypeptide is selected from the group consisting of an antibody, a Fab, an F(ab′)2, an Fv, and SCA and a CDR.
- 33. The reaction substrate of claim 29, wherein each capture probe is a first oligonucleotide, each capture probe specific domain is a second oligonucleotide, and each target analyte specific domain is as hapten binding polypeptide.
- 34. The reaction substrate of claim 29, wherein each capture probe is a hapten binding polypeptide, each capture probe specific domain is a hapten, and each target analyte specific domain is an oligonucleotide.
- 35. The reaction substrate of claim 34, wherein each hapten binding polypeptide is selected from the group consisting of an antibody, an FV, and an Fab.
- 36. The reaction substrate of claim 29, wherein each capture probe is an avidin, each capture probe specific domain is a biotin, and each target analyte specific domain is an oligonucleotide.
- 37. The reaction substrate of claim 30, 31, 35, or 36, wherein each target analyte specific domain is an oligonucleotide nucleic acid amplification primer.
- 38. The reaction substrate of claim 25, wherein at least one reaction substrate is contained in at least one reaction chamber.
- 39. The reaction substrate of claim 38, wherein a plurality of reaction substrates are contained within each reaction chamber.
- 40. The reaction substrate of claim 40, further including a reaction vessel, wherein a plurality of reaction chambers are contained within said reaction vessel.
- 41. The reaction substrate of claim 39, wherein said reaction vessel comprises about 2 to about 10,000 reaction chambers.
- 42. The reaction substrate of claim 25, wherein said reaction substrate is optically clear.
- 43. The reaction substrate of claim 25, wherein each capture probe has a percentage base composition in the range of about 30-40% G, 30-40% C, 10-20% A, and 10-20% T.
- 44. The set of claim 25, wherein each capture probe has a length ranging from 2 to 30 bases.
- 45. The set of claim 44, wherein each capture probe has a length ranging from 5 to 25 bases.
- 46. The set of claim 45, wherein each capture probe has a length ranging from 10 to 20 bases.
- 47. The set of claim 25, wherein each capture probe has a length of about 16 bases.
- 48. The set of claim 25, wherein each capture probes has a length that differs by no more than one base from the average length of all capture probes.
- 49. The set of claim 25, wherein each capture probe possesses an overall gross base composition which is substantially similar to the overall gross base composition of all other capture probes.
- 50. The set of claim 25, wherein each capture probe has a sequence homology that differs from the sequence homology of each other capture probe by at least 20%.
- 51. The set of claim 50, wherein each capture probe has a sequence homology that differs from the sequence homology of each other capture probe by at least 40%.
- 52. The set of claim 51, wherein each capture probe has a sequence homology that differs from the sequence homology of each other capture probe by at least 50%.
- 53. The set of claim 52, wherein each capture probe has a sequence homology that differs from the sequence homology of each other capture probe by at least 60%.
- 54. The set of claim 25, wherein the sequence homology of any two capture probes is less than 80%.
- 55. A reaction substrate for use in conducting multiplexed microassays to determine binding between a target molecule and a capture probe, said reaction substrate having an array of parallel printed biosites, wherein each biosite comprises a single type of capture probe bound to said reaction substrate.
- 56. A method for preparing a reaction substrate for use as an assay device, comprising the step of parallel printing an array of biosites on said reaction substrate, wherein each biosite comprises a single type of capture probe bound to said reaction substrate.
- 57. The method of claim 56, further including the step of binding a plurality of target probes to said array of biosites, wherein each target probe binds to a specific capture probe within said array.
- 58. The method of claim 56, wherein each biosite comprises a spot of about 25 to about 200 μm in diameter.
- 59. The method of claim 56, wherein said reaction substrate is optically clear and has a thickness of about 50 to about 300 μm.
- 60. A method for identifying target analytes in a plurality of separate samples, each target analyte capable of binding a corresponding capture probe, including the steps of:
parallel printing an array of biosites on a reaction substrate, each biosite comprising a single type of capture probe bound to said substrate; contacting each biosite with a sample including at least one target analyte; and determining the presence or absence of binding of a target analyte at each biosite in said array.
- 61. The method of claim 60, wherein each target analyte is detectably labeled.
- 62. The method of claim 61, wherein each target analyte is labeled with a fluorescent label.
- 63. The method of claim 61, wherein each target analyte is labeled with an electroluminescent label.
- 64. The method of claim 61, wherein each target analyte is labeled with a radioisotope label.
- 65. The method of claim 60, wherein the presence or absence of binding at each biosite is determined using an optical sensing array.
- 66. The method of claim 65, wherein the optical sensing array is disposed in close proximity to the reaction substrate.
- 67. The method of claim 65, wherein the optical sensing array is lensless.
- 68. An apparatus for preparing a reaction substrate for use as an assay device, including:
an attachment site for holding a plurality of flexible capillary tubes at a point spaced from the distal ends of such capillary tubes; a structure for positioning the proximal end of each capillary tube within a corresponding supply chamber, each supply chamber capable of supplying a liquid reagent to at least one corresponding capillary tube; a print head for holding said plurality of capillary tubes near their distal ends and for precisely positioning the distal ends of said capillary tubes with respect to said reaction substrate to deposit liquid reagents onto said reaction substrate as biosites.
- 69. The apparatus of claim 68, where the print head includes an ink-jet deposition device.
- 70. A method for detecting labeled sample molecules, comprising the steps of:
providing a reaction substrate having a plurality of biosites, each biosite being attached to the reaction substrate, the reaction substrate having an embedded conductive material connected to an electrical source; contacting each biosite with a sample of labeled molecules; initiating an electrochemical event within the labeled molecules which releases electromagnetic energy; detecting the released electromagnetic energy.
- 71. The method of claim 70, wherein the sample molecules are fluorescently labeled.
- 72. The method of claim 70, wherein the sample molecules are electroluminescently labeled.
- 73. The method of claim 70, wherein the released electromagnetic energy is detected using an optical sensing array.
- 74. The method of claim 73, wherein the optical sensing array is disposed in close proximity to the reaction substrate.
- 75. The method of claim 73, wherein the optical sensing array is lensless.
- 76. A method for detecting particle emissions from labeled sample molecules, comprising the steps of:
providing a reaction substrate having a plurality of biosites, each biosite comprising a single type of capture probe bound to said substrate; providing a lensless array of particle detectors disposed in close proximity to said plurality of biosites; contacting each biosite with a sample of labeled molecules; converting, in said particle detectors, particle emissions emanating directly from said sample into corresponding electrical signals; forming an image from said electrical signals representing the quantitative presence or absence of the labeled molecules on each biosite from said sample.
- 77. The method of claim 76, wherein the sample molecules are fluorescently labeled.
- 78. The method of claim 76, wherein the sample molecules are chemiluminescently labeled.
- 79. The method of claim 76, wherein the sample molecules are radioisotope labeled.
- 80. A set of nucleic acid capture probes, each configured to be bound to a reaction substrate, each capture probe having a length sufficient to provide dissimilarity among capture probes, each capture probe having a percentage base composition in the range of about 30-40% G, 30-40% C, 10-20% A, and 10-20% T, wherein the set is selected such that the nucleic acid sequence of each capture probe in the set is substantially dissimilar from the nucleic acid sequence of all other capture probes in the set.
- 81. The set of claim 80, wherein the length ranges from 2 to 30 bases.
- 82. The set of claim 81, wherein the length ranges from 5 to 25 bases.
- 83. The set of claim 82, wherein the length ranges from 10 to 20 bases.
- 84. The set of claim 80, wherein the length is about 16 bases.
- 85. The set of claim 80, wherein the lengths of said capture probes differ by no more than one base from the average length of the capture probes.
- 86. The set of claim 80, wherein each capture probe possesses an overall gross base composition which is substantially similar to the overall gross base composition of all other capture probes of the set.
- 87. The set of claim 80, wherein each capture probe of a given binding specificity has a nucleic acid sequence that differs from the nucleic acid sequence of each other capture probe of different specificity by at least 20%.
- 88. The set of claim 87, wherein each capture probe of a given binding specificity has a nucleic acid sequence that differs from the nucleic acid sequence of each other capture probe of different binding specificity by at least 40%.
- 89. The set of claim 88, wherein each capture probe of a given binding specificity has a nucleic acid sequence that differs from the nucleic acid sequence of each other capture probe of different binding specificity by at least 50%.
- 90. The set of claim 89, wherein each capture probe of a given binding specificity has a nucleic acid sequence that differs from the nucleic acid sequence of each other capture probe of different binding specificity by at least 60%.
- 91. The set of claim 90, wherein each capture probe of a given binding specificity has a nucleic acid sequence that differs from the nucleic acid sequence of each other capture probe of different binding specificity by at least 80%.
- 92. The set of claim 80, wherein the set includes at least 16 capture probes.
Parent Case Info
[0001] This application is based on U.S. provisional application 60/034,627, filed Dec. 31, 1996, incorporated herein by reference.
STATEMENT AS TO FEDERALLY SPONSORED RESEARCH
[0002] This invention was made at least in part with funds from the National Aeronautics and Space Administration, Grant Number NAGW 4530.
Divisions (2)
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Number |
Date |
Country |
Parent |
09217154 |
Dec 1998 |
US |
Child |
10316077 |
Dec 2002 |
US |
Parent |
09002170 |
Dec 1997 |
US |
Child |
10316077 |
Dec 2002 |
US |
Continuations (1)
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Number |
Date |
Country |
Parent |
09625086 |
Jul 2000 |
US |
Child |
10316077 |
Dec 2002 |
US |