Ultra-sensitive detection of molecules or particles using beads or other capture objects
First Claim
1. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:
- providing a substrate comprising a plurality of locations, at least a portion of which locations contain a bead, wherein with respect to the total number of beads present on the substrate, the ratio of beads comprising at least one analyte molecule or particle to beads comprising no analyte molecules or particles is between about 8;
1 and about 1;
10,000,000;
addressing at least a portion of the plurality of locations, wherein during the addressing step at least two of the plurality of locations is addressed at least partially concurrently;
detecting at each addressed location the presence or absence of a bead and whether, if present, the bead comprises any analyte molecules or particles; and
determining a measure of the concentration of analyte molecules or particles in the fluid sample at least in part by determining the number of locations addressed containing a bead comprising at least one analyte molecule or particle.
1 Assignment
0 Petitions
Accused Products
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 with respect to a plurality of capture objects. At least a portion of the plurality of capture objects may be spatially separated into a plurality of locations. A measure of the concentration of analyte molecules in a fluid sample may be determined, at least in part, on the number of reaction vessels comprising an analyte molecule immobilized with respect to a capture object. In some cases, the assay may additionally comprise steps including binding ligands, precursor labeling agents, and/or enzymatic components.
-
Citations
39 Claims
-
1. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:
-
providing a substrate comprising a plurality of locations, at least a portion of which locations contain a bead, wherein with respect to the total number of beads present on the substrate, the ratio of beads comprising at least one analyte molecule or particle to beads comprising no analyte molecules or particles is between about 8;
1 and about 1;
10,000,000;addressing at least a portion of the plurality of locations, wherein during the addressing step at least two of the plurality of locations is addressed at least partially concurrently; detecting at each addressed location the presence or absence of a bead and whether, if present, the bead comprises any analyte molecules or particles; and determining a measure of the concentration of analyte molecules or particles in the fluid sample at least in part by determining the number of locations addressed containing a bead comprising at least one analyte molecule or particle. - View Dependent Claims (5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 22, 23)
-
-
2. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:
-
providing a substrate comprising a plurality of locations, at least a portion of which contain a bead, wherein with respect to the total number of beads present on the substrate, the ratio of beads comprising at least one analyte molecule or particle associated with a binding ligand to beads comprising no analyte molecules or particles associated with a binding ligand is between about 8;
1 and about 1;
10,000,000;addressing at least a portion of the plurality of locations, wherein during the addressing step at least two of the plurality of locations is addressed at least partially concurrently; detecting at each addressed location the presence or absence of a bead and whether, if present, the bead comprises any analyte molecules or particles associated with a binding ligand; and determining a measure of the concentration of analyte molecules or particles in the fluid sample at least in part by determining the number of locations addressed containing a bead comprising at least one analyte molecule or particle associated with a binding ligand. - View Dependent Claims (19, 20, 21)
-
-
3. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:
-
immobilizing a plurality of analyte molecules or particles with respect to a plurality of beads; spatially segregating at least a portion of the plurality of beads into a plurality of separate locations; addressing at least some of the plurality of locations and determining the number of locations containing a bead; further determining the number of said locations containing a bead and an analyte molecule or particle; and determining a measure of the concentration of analyte molecules or particles in the fluid sample based at least in part on the ratio of the number of locations containing a bead and an analyte molecule and particle, to the number of locations containing a bead. - View Dependent Claims (24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
-
-
4. A method for determining a measure of the concentration of analyte molecules or particles in a fluid sample, comprising:
-
immobilizing a plurality of analyte molecules or particles with respect to a plurality of beads; spatially segregating at least a portion of the plurality of beads into a plurality of separate locations; addressing at least some of the plurality of locations and determining the number of locations containing a bead; further determining the number of said locations containing a bead and an analyte molecule or particle; and determining a measure of the concentration of analyte molecules or particles in the fluid sample based at least in part on the ratio of the number of locations containing a bead and an analyte molecule and particle, to the number of locations containing a bead but not containing any analyte molecules or particles.
-
Specification