Ultra-sensitive detection of molecules on single molecule arrays
First Claim
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1. A method of detecting analyte molecules or particles in a fluid sample, comprising:
- exposing a substrate comprising a plurality of capture components to a sample comprising a plurality of analyte molecules or particles, so that analyte molecules or particles associate with capture components to form a plurality of complexes, each complex being immobilized with respect to the substrate and comprising at least one capture component and at least one analyte molecule or particle;
dissociating at least a portion of each complex to form a plurality of dissociated species, which are not immobilized with respect to the substrate;
partitioning the plurality of dissociated species across a plurality of reaction vessels;
determining the presence or absence of a dissociated species in at least one reaction vessel; and
determining the number of the plurality of reaction vessels and/or fraction of the plurality of reaction vessels that contain or do not contain a dissociated species,wherein the plurality of dissociated species are partitioned such that a statistically significant fraction of the reaction vessels contain no dissociated species and a statistically significant fraction of reaction vessels contain at least one dissociated species.
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Abstract
The present invention relates to systems and methods for detecting analyte molecules or particles in a fluid sample and in some cases, determining a measure of the concentration of the molecules or particles in the fluid sample. Methods of the present invention may comprise immobilizing a plurality of analyte molecules or particles to form a plurality of complexes, releasing at least a portion of some of the plurality of complexes, determining at least a portion of the plurality of complexes released, and determining a measure of the concentration of the analyte molecules or particles in a fluid sample.
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39 Claims
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1. A method of detecting analyte molecules or particles in a fluid sample, comprising:
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exposing a substrate comprising a plurality of capture components to a sample comprising a plurality of analyte molecules or particles, so that analyte molecules or particles associate with capture components to form a plurality of complexes, each complex being immobilized with respect to the substrate and comprising at least one capture component and at least one analyte molecule or particle; dissociating at least a portion of each complex to form a plurality of dissociated species, which are not immobilized with respect to the substrate; partitioning the plurality of dissociated species across a plurality of reaction vessels; determining the presence or absence of a dissociated species in at least one reaction vessel; and determining the number of the plurality of reaction vessels and/or fraction of the plurality of reaction vessels that contain or do not contain a dissociated species, wherein the plurality of dissociated species are partitioned such that a statistically significant fraction of the reaction vessels contain no dissociated species and a statistically significant fraction of reaction vessels contain at least one dissociated species. - 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)
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27. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:
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capturing a plurality of analyte molecules or particles on a first substrate; releasing a plurality of molecules or particles from the first substrate; detecting molecules or particles released from the first substrate on or within a second substrate comprising a plurality of reaction vessels; and determining a measure of the concentration of the analyte molecules or particles in the fluid sample based on the detection of molecules or particles released from the first substrate on or within the second substrate, wherein the measure of the concentration of the analyte molecules or particles in the fluid sample is determined by determining the number or fraction of the plurality of reaction vessels that contain or do not contain a molecule or particle released from the first substrate. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
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Specification