US 12,338,487 B2
Multiomic analysis of cell analytes using microfluidic systems
Harold G. Craighead, Ithaca, NY (US); Harvey C. Tian, Ithaca, NY (US); David M. Lin, Ithaca, NY (US); and Adam J. Bisogni, Ithaca, NY (US)
Assigned to CORNELL UNIVERSITY, Ithaca, NY (US)
Appl. No. 17/298,533
Filed by CORNELL UNIVERSITY, Ithaca, NY (US)
PCT Filed Nov. 29, 2019, PCT No. PCT/US2019/063866
§ 371(c)(1), (2) Date May 28, 2021,
PCT Pub. No. WO2020/113173, PCT Pub. Date Jun. 4, 2020.
Claims priority of provisional application 62/772,621, filed on Nov. 28, 2018.
Prior Publication US 2022/0010360 A1, Jan. 13, 2022
Int. Cl. C12Q 1/00 (2006.01); B01L 3/00 (2006.01); C12N 15/10 (2006.01); C12Q 1/6806 (2018.01); C12M 3/06 (2006.01)
CPC C12Q 1/6806 (2013.01) [B01L 3/5027 (2013.01); B01L 3/502761 (2013.01); C12N 15/1006 (2013.01); B01L 2200/0668 (2013.01); B01L 2300/0816 (2013.01); B01L 2400/086 (2013.01); C12M 23/16 (2013.01)] 23 Claims
OG exemplary drawing
 
1. A method for isolating one or more distinct analyte component from a cell sample, said method comprising the acts of:
introducing, under continuous flow conditions, a cell sample comprising at least one cell into a microfluidic device comprising an array of cell capture micropillars and a plurality of nucleic acid entanglement micropillars
capturing the at least one cell in the array of cell capture micropillars while the microfluidic device is subjected to a continuous flow rate;
treating the at least one captured cell with a sequential workflow procedure under conditions effective to separate at least one analyte therefrom, wherein said analyte is selected from the group consisting of: (i) a total protein fraction; (ii) a plasma membrane protein fraction; (iii) a total RNA fraction; (iv) a cytosolic RNA fraction; (v) a cytosolic protein fraction; (vi) a nuclear RNA fraction; (vii) a nuclear protein fraction; (viii) a chromatin fraction comprising genomic DNA (gDNA) regions of open chromatin; (ix) a gDNA markers fraction comprising epigenetic and regulatory markers of gDNA; (x) an amplified gDNA fraction; and (xi) a methylated gDNA fraction, wherein said gDNA is single-stranded gDNA, double-stranded gDNA, or a combination of single-and double-stranded gDNA; and
isolating the analyte in a manner suitable for further testing and/or analysis thereof.