US 11,789,016 B2
Methods, systems and kits for in-pen assays
Troy A. Lionberger, Berkeley, CA (US); Phillip J. M. Elms, Oakland, CA (US); Anupam Singhal, Berkeley, CA (US); Randall D. Lowe, Jr., Emeryville, CA (US); Volker L. S. Kurz, Oakland, CA (US); and Paul M. Lebel, Redwood City, CA (US)
Assigned to PHENOMEX INC., Emeryville, CA (US)
Filed by Berkeley Lights, Inc., Emeryville, CA (US)
Filed on Apr. 15, 2020, as Appl. No. 16/849,811.
Application 16/849,811 is a continuation of application No. PCT/US2018/055918, filed on Oct. 15, 2018.
Claims priority of provisional application 62/572,525, filed on Oct. 15, 2017.
Prior Publication US 2020/0408751 A1, Dec. 31, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. B01L 3/00 (2006.01); G01N 33/543 (2006.01); G01N 33/68 (2006.01)
CPC G01N 33/54366 (2013.01) [B01L 3/502761 (2013.01); G01N 33/6854 (2013.01); B01L 2200/0668 (2013.01); B01L 2300/0654 (2013.01); B01L 2300/0864 (2013.01); B01L 2300/165 (2013.01); B01L 2300/1822 (2013.01); B01L 2400/0424 (2013.01); B01L 2400/0457 (2013.01); B01L 2400/0472 (2013.01); B01L 2400/086 (2013.01)] 23 Claims
OG exemplary drawing
 
1. A method of assessing a level of secretion of an analyte by a biological micro-object, or a population of biological micro-objects generated therefrom, the method comprising:
introducing the biological micro-object into a sequestration pen of a microfluidic device, wherein the microfluidic device comprises an enclosure having a flow region, wherein the sequestration pen is fluidically connected to the flow region, and wherein the sequestration pen contains a first fluidic medium;
allowing the biological micro-object, or the population of biological micro-objects generated therefrom, to secrete an analyte comprising an exogenous tag into the first fluidic medium within the sequestration pen;
introducing a second fluidic medium into the flow region, wherein the second fluidic medium comprises a plurality of reporter molecules, and wherein each reporter molecule comprises:
a binding component configured to bind the exogenous tag of the secreted analyte; and
a detectable label;
allowing a portion of the plurality of reporter molecules to diffuse into the sequestration pen and bind to the analyte secreted therein, thereby producing a plurality of reporter molecule: secreted analyte (RMSA) complexes; and
detecting reporter molecules located within an area of interest within the microfluidic device, wherein the area of interest includes at least a portion of the sequestration pen.