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US 2024/0200134 A1Published application
Single cell analyses
AssigneeMassachusetts Institute of Technology
InventorsJ. Christopher Love, Todd Michael Gierahn, Alexander K. Shalek, Marc Havens Wadsworth, Travis K. Hughes
Life of this patent
Estimating the term…
Abstract
This disclosure describes improvements to both hardware and enzymatic reactions used in single cell analyses such as but not limited to Seq-well that enable significant increases in the yield of transcripts per cell, improved portability and case of use, increased scalability of the assay, and linkage of transcript information to other measurements made in the picowell arrays.
Claims17
1-119. (canceled)
A microwell device comprising:
- (a) a photoresist film comprising a top surface, a bottom surface, and a plurality of through-holes from the top to the bottom surface, wherein each hole has a top opening on the top surface and a bottom opening on the bottom surface, and
- (b) a porous bottom membrane in contact with the bottom surface of the photoresist film, wherein the bottom membrane has a flux rate of at least 0.1 mL/min/cm2 as measured by the initial flux rate of water at 10 pounds per square inch (psi).
122-126. (canceled)
A method of cellular loading comprising,
- (a) flowing a fluid sample comprising a plurality of cells through a microarray device, said device comprising, (i) a bottomless microwell array comprising a top surface, a bottom surface, and a plurality of through-holes from the top to the bottom surface, wherein each hole has a top opening on the top surface and a bottom opening on the bottom surface, and (ii) a porous bottom membrane in contact with the bottom surface of the microwell array, wherein the bottom membrane has a flux rate of at least 0.1 mL/min/cm2 as measured by the initial flux rate of water at 10 pounds per square inch (psi),
- (b) applying a pressure gradient from the top opening to the bottom opening of at least one of the plurality of through-holes, thereby loading a cell into said at least one through-hole, and
- (c) retaining the cell at the bottom of the at least one through-hole by applying the pressure gradient, thereby loading cells in said through-holes in the microarray.
(canceled)
133-134. (canceled)
Dependencies are inferred from the claim text.