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US 2017/0283859 A1Published application
MICROFLUIDIC qRT-PCR ANALYSIS OF SINGLE CELLS
AssigneeThe Trustees of Columbia University in the City of New York
InventorsQiao Lin, Jing Zhu, Timothy R. Olsen, Hao Sun, Brian Ponnaiya, Sally A. Amundson, David J. Brenner
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Abstract
The disclosed subject matter provides a microdevice and techniques for single-cell gene expression profiling using a microfluidic device capable of cell-trapping, cell lysis, bead-based gene analysis. The microdevice can be capable of independent or parallelized, simultaneous quantitative genetic assays of single cells.
Claims23
A microfluidic device for single cell gene expression analysis comprising:
- a cell inlet configured to receive a fluid containing a plurality of cells;
- and
- one or more analysis units coupled to the cell inlet, wherein each of the one or more analysis units comprise:
- a cell trap configured to trap a single cell from the plurality of cells;
- a reaction chamber coupled to the cell trap;
- one or more magnetic microbeads comprising a primer configured to capture mRNA obtained by lysing the single cell;
- and
- one or more magnets configured to transport the one or more microbeads between the cell trap and the reaction chamber.
5 more elements
A method for single cell gene expression analysis comprising:
- (a) providing a microfluidic device comprising:
- a cell inlet and one or more analysis units coupled to the cell inlet, wherein each of the one or more analysis units comprise a cell trap, a reaction chamber coupled to the cell trap, one or more magnetic microbeads comprising at least one primer configured to capture cell mRNA, and one or more magnets;
- (b) optionally isolating one or more analysis units from the cell inlet;
- (c) trapping a single cell in the cell trap of at least one analysis unit;
- (d) removing excess cells from the device;
- (e) lysing the single cell with a lysis buffer;
- (f) capturing mRNA obtained by lysing the single cell with the one or more magnetic microbeads comprising at least one primer;
- (g) transferring the one or more magnetic microbeads to the reaction chamber of the selected analysis unit for gene expression analysis reactions using the one or more magnets.
5 more elements
Dependencies are inferred from the claim text.