Apparatus and method for analysis of molecules
First Claim
Patent Images
1. An apparatus comprising:
- an array of optical confinements having a density on a substrate exceeding 4×
104 confinements per mm2, wherein said optical confinements are suitable for holding a biological reagent, and wherein said optical confinements provide an effective observation volume that permits observation of individual molecules present in said biological reagent; and
an optical system operatively coupled to the optical confinements that detects signals from the effective observation volume of said confinements.
2 Assignments
0 Petitions
Accused Products
Abstract
The present invention relates to optical confinements, methods of preparing and methods of using them for analyzing molecules and/or monitoring chemical reactions. The apparatus and methods embodied in the present invention are particularly useful for high-throughput and low-cost single-molecular analysis.
-
Citations
48 Claims
-
1. An apparatus comprising:
-
an array of optical confinements having a density on a substrate exceeding 4×
104 confinements per mm2, wherein said optical confinements are suitable for holding a biological reagent, and wherein said optical confinements provide an effective observation volume that permits observation of individual molecules present in said biological reagent; and
an optical system operatively coupled to the optical confinements that detects signals from the effective observation volume of said confinements. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 41, 42, 43, 44)
-
-
36. A method of detecting a biological analyte, comprising:
-
optically capturing the analyte within an optical confinement that is created by (a) providing an array of optical confinements having a density on a substrate exceeding 4×
104 confinements per mm2, wherein said optical confinements provide an effective observation volume that permits observation of individual molecules; and
an optical system operatively coupled to the optical confinements that detects signals from the effective observation volume of said confinements; and
(b) illuminating at least one optical confinement within the array that is suspected to contain the analyte with an incident light beam thereby detecting the analyte. - View Dependent Claims (37, 38, 39)
-
-
45. A method of sequencing a plurality of target nucleic acid molecules, comprising:
-
(a) providing an array of optical confinements having a density on a substrate exceeding 4×
104 confinements per mm2, wherein said optical confinements provide an effective observation volume that permits observation of individual molecules; and
an optical system operatively coupled to the optical confinements that detects signals from the effective observation volume of said confinement;
(b) mixing in the optical confinements the plurality of target nucleic acid molecules, primers complementary to the target nucleic acid molecules, polymerization enzymes, and more than one type of nucleotides or nucleotide analogs to be incorporated into a plurality of nascent nucleotide strands, each strand being complementary to a respective target nucleic acid molecule;
(c) subjecting the mixture of step (b) to a polymerization reaction under conditions suitable for formation of the nascent nucleotide strands by template-directed polymerization of the nucleotides or nucleotide analogs;
(d) illuminating the optical confinements with an incident light beam; and
(e) identifying the nucleotides or the nucleotide analogs incorporated into the each nascent nucleotide strand. - View Dependent Claims (46, 47, 48)
-
Specification