Methods and arrays for detecting cells and cellular components in small defined volumes
First Claim
Patent Images
1. A method of producing an array of single cells, the method comprising:
- a) providing a sample and an array, said sample comprising cells, said array comprising at least 1000 sites, each site having a defined volume between 10 attoliters and 50 picoliters; and
b) contacting said array with said sample such that the ratio of cells to each site of said array is less than 1;
1, so as to produce an array comprising sites containing single molecules.
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Abstract
Arrays of single cells and methods of producing an array of single cells are described. Arrays with defined volumes between 10 attoliters and 50 picoliters enable single cell capture, detection and quantitation.
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Citations
53 Claims
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1. A method of producing an array of single cells, the method comprising:
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a) providing a sample and an array, said sample comprising cells, said array comprising at least 1000 sites, each site having a defined volume between 10 attoliters and 50 picoliters; and
b) contacting said array with said sample such that the ratio of cells to each site of said array is less than 1;
1, so as to produce an array comprising sites containing single molecules. - 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)
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24. A method of detecting cells in a sample, the method comprising:
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a) providing a sample and an array, said sample comprising cells in a first concentration, said array comprising at least 1000 sites, each site having a defined volume between 10 attoliters and 50 picoliters;
b) diluting said sample to create a diluted sample, said diluted sample comprising cells in a second concentration;
c) contacting said array with said diluted sample such that the ratio of cells to each site of said array is less than 1;
1; and
d) determining the number of sites of said array which contain a single cell. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36)
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37. A method of detecting a target analyte of a cell in a sample, the method comprising:
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a) providing a sample and an array, said sample comprising cells in a first concentration, each cell comprising a target analyte, said array comprising at least 1000 sites, each site having a defined femtoliter volume;
b) diluting said sample to create a diluted sample, said diluted sample comprising cells in a second concentration;
c) contacting said array with said diluted sample such that the ratio of cells to each site of said array is less than 1;
1;
d) treating said cells in said array under conditions such that said cells are lysed; and
e) determining the number of sites of said array which contain a target analyte of said cells. - View Dependent Claims (38, 39, 40, 41, 42, 43, 44)
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45. A method of detecting a target analyte of a cell in a sample, the method comprising:
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a) providing a sample and an array, said sample comprising cells in a concentration, said array comprising at least 1000 sites, each site having a defined femtoliter volume;
b) treating said cells under conditions such that said cells are lysed so as to create a treated sample;
c) contacting said array with said treated sample such that the ratio of target analyte to each site of said array is less than 1;
1; and
d) determining the number of sites of said array which contain a target analyte. - View Dependent Claims (46, 47, 48, 49, 50, 51, 52, 53)
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Specification