Method for analyzing molecular expression or function in an intact single cell
First Claim
1. A method of isolating and harvesting a substantially intact single cell from its organ tissue comprising the following steps:
- (i) subjecting a tissue mass to a dissociation method so that the cells are dissociated from the tissue to expose cell bodies or cell processes;
(ii) contacting a dissociated cell on its membrane with a device capable of collecting the cell substantially intact from said tissue;
(iii) withdrawing said device with the cell attached; and
(iv) harvesting said cell, wherein said cell is isolated intact from its organ tissue.
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Abstract
A method for obtaining molecules from a substantially intact single cell is disclosed. The method differs from prior methods that obtain a variable fraction of a cell'"'"'s contents and therefore cannot quantitatively estimate the number of molecules in the cells. The present method comprises isolating and harvesting a substantially intact single cell from its organ tissue comprising the steps of subjecting a tissue mass to a dissociation method so that the cells are dissociated from the tissue to expose cell bodies or cell processes, contacting a dissociated cell with a device capable of collecting the cell from the tissue substantially intact, withdrawing device with the cell attached, and then isolating or detecting the molecules in the single cell.
31 Citations
40 Claims
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1. A method of isolating and harvesting a substantially intact single cell from its organ tissue comprising the following steps:
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(i) subjecting a tissue mass to a dissociation method so that the cells are dissociated from the tissue to expose cell bodies or cell processes;
(ii) contacting a dissociated cell on its membrane with a device capable of collecting the cell substantially intact from said tissue;
(iii) withdrawing said device with the cell attached; and
(iv) harvesting said cell, wherein said cell is isolated intact from its organ tissue. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A method for obtaining molecules from a single cell comprising:
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(i) isolating a substantially intact single cell from its organ tissue comprising the following steps;
(a) subjecting a tissue mass to a dissociation method so that the cells are dissociated from the tissue to expose cell bodies or cell processes;
(b) contacting a dissociated cell on its membrane with a device capable of collecting the cell substantially intact from said tissue;
(c) withdrawing said device with the cell attached; and
(ii) isolating or detecting said molecules in the single cell. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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27. A method for measuring a molecule(s) from a single cell comprising:
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(i) isolating and harvesting a substantially intact single cell from its organ tissue comprising the following steps;
(a) subjecting a tissue mass to a dissociation method so that the cells are dissociated from the tissue to expose cell bodies or cell processes;
(b) contacting a dissociated cell on its membrane with a device capable of collecting the cell substantially intact from said tissue;
(c) withdrawing said device with the cell attached;
(d) harvesting said cell, wherein said cell is isolated intact from its organ tissue; and
(ii) measuring said molecule(s) in the single cell. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
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Specification
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- Resources
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Current AssigneeUniversity Of Kentucky Research Foundation (University of Kentucky)
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Original AssigneeUniversity Of Kentucky Research Foundation (University of Kentucky)
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InventorsBlalock, Eric, Chen, Kuey-Chu, Kaminker, Patrick, Landfield, Philip W., Thibault, Olivier
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Primary Examiner(s)Horlick, Kenneth R.
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Assistant Examiner(s)Spiegler, Alexander H.
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Application NumberUS09/627,082Time in Patent Office719 DaysField of Search435/4, 435/1.1, 435/6, 435/91.1, 435/91.2, 435/270US Class Current435/4CPC Class CodesG01N 33/5005 involving human or animal c...
