US 12,011,719 B2
Digital microfluidic system for single-cell isolation and characterization of analytes
Arash Jamshidi, San Francisco, CA (US); Yan-you Lin, San Francisco, CA (US); Farnaz Absalan, San Francisco, CA (US); Sarah Stuart, San Francisco, CA (US); Gordon Cann, San Francisco, CA (US); Yir-Shyuan Wu, San Francisco, CA (US); Tarun Khurana, San Francisco, CA (US); and Jeffrey S Fisher, San Francisco, CA (US)
Assigned to Illumina, Inc., San Diego, CA (US)
Filed by Illumina, Inc., San Diego, CA (US)
Filed on Dec. 17, 2021, as Appl. No. 17/555,013.
Application 17/555,013 is a division of application No. 15/779,459, granted, now 11,203,016, previously published as PCT/US2016/064206, filed on Nov. 30, 2016.
Claims priority of provisional application 62/358,968, filed on Jul. 6, 2016.
Claims priority of provisional application 62/314,071, filed on Mar. 28, 2016.
Claims priority of provisional application 62/281,510, filed on Jan. 21, 2016.
Claims priority of provisional application 62/261,786, filed on Dec. 1, 2015.
Prior Publication US 2022/0184622 A1, Jun. 16, 2022
Int. Cl. B01L 3/00 (2006.01); C12Q 1/6806 (2018.01); G01N 1/28 (2006.01); G01N 33/543 (2006.01); C12Q 1/6869 (2018.01); G01N 15/10 (2006.01)
CPC B01L 3/502761 (2013.01) [B01L 3/502784 (2013.01); B01L 3/5088 (2013.01); C12Q 1/6806 (2013.01); G01N 1/28 (2013.01); G01N 33/54333 (2013.01); B01L 2200/0605 (2013.01); B01L 2200/0642 (2013.01); B01L 2200/0668 (2013.01); B01L 2300/0819 (2013.01); B01L 2300/165 (2013.01); B01L 2400/0427 (2013.01); B01L 2400/043 (2013.01); C12Q 1/6869 (2013.01); C12Q 2563/143 (2013.01); C12Q 2563/149 (2013.01); C12Q 2563/185 (2013.01); G01N 2015/1006 (2013.01)] 9 Claims
 
1. A method comprising:
(a) utilizing a droplet actuator to transport a sample droplet to a microwell device, the sample droplet including cells of interest, the microwell device including
a bottom substrate having a plurality of microwells that open onto a droplet operations surface of the microwell device, and
a top substrate comprising a plurality of molded cell traps that extend into the droplet operations surface of the microwell device and are located above each of the plurality of microwells, wherein the plurality of microwells and the plurality of molded cell traps are configured such that the cells of interest are trapped by the plurality of molded cell traps and are thereby deposited into the plurality of microwells;
(b) introducing capture beads to the microwells, wherein capture elements are immobilized on the capture beads; and
(c) utilizing the droplet actuator to transport a cell lysis reagent droplet to the microwell device, wherein portions of the cell lysis reagent droplet enter the microwells and, during an incubation period, cause the cells of interest to lyse and release analyte that is captured by the capture elements on the capture beads.