In situ-generated microfluidic assay structures, related kits, and methods of use thereof
AssigneeBerkeley Lights, Inc.
InventorsKristin G. Beaumont, Peter J. Beemiller, Volker L. S. Kurz, Gregory G. Lavieu, Xiaohua Wang, Aathavan Karunakaran
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Abstract
In situ-generated microfluidic capture structures incorporating a solidified polymer network, methods of preparation and use, compositions and kits therefor are described. Microfluidic capture structures may be advantageously used for assays performed within the microfluidic environment, providing flexibility in assaying micro-objects such as biological cells. Assay reagents and analytes may be incorporated within the microfluidic capture structures.
Claims29
A microfluidic device comprising:
- an enclosure comprising a substrate and microfluidic circuit material, the enclosure defining a flow region and at least one sequestration pen, each located within the enclosure;
- and
- at least one in situ-generated capture structure disposed within the at least one sequestration pen, wherein the at least one in situ-generated capture structure comprises a solidified polymer network, and wherein the solidified polymer network comprises one or more functionalized sites.
A method of assaying a micro-object in a microfluidic device comprising at least a first in situ-generated capture structure comprising:
- disposing a micro-object within the microfluidic device in a region proximal to the at least first in situ-generated capture structure comprising a solidified polymer network, wherein the solidified polymer network comprises an assay reagent or assay analyte;
- contacting the assay reagent or assay analyte with the micro-object or a biological product of the micro-object;
- and
- detecting an interaction of the assay reagent or assay analyte with the micro-object or the biological product.
A method of preparing a microfluidic device comprising at least a first in situ-generated capture structure, the method comprising:
- providing the microfluidic device, wherein the microfluidic device comprises an enclosure comprising a substrate and microfluidic circuit material, the enclosure defining a flow region and at least one sequestration pen fluidically connected to the flow region;
- introducing a first flowable functionalized pre-polymer into the flow region;
- and
- activating solidification of the first flowable functionalized pre-polymer at at least one selected area of the at least one sequestration pen, thereby forming the at least a first in situ-generated capture structure therein.
Dependencies are inferred from the claim text.