US 12,650,367 B2 · Systems and methods for encapsulation and multi-step processing of biological samples

Abstract

This invention relates to methods and systems for isolation of species in semi-permeable capsules and processing of encapsulated species through series of steps and/or reactions. To produce capsules, first aqueous two-phase system (ATPS) droplets are generated using microfluidics system. Then the hydrogel shell layer is hardened by inducing polymerization. As exemplified in this invention to achieve concentric ATPS droplet formation density-matched PEGDA and Dextran polymer solutions can be used. Once a capsule is formed, its composition can be changed by adding new reagents or replacing out old ones (e.g. by resuspending capsules in desired aqueous solution). The hydrogel shell of semi-permeable capsules can be dissolved at selected step during multi-step procedures to release the encapsulated species. This invention exemplifies isolation of individual cells within capsules and using the encapsulated cells for genotypic and phenotypic analysis. This invention also exemplifies use of capsules in multi-step procedures to perform complex biological reactions.

Claims30

Dependent claims

A method of performing incompatible reactions in a common reaction volume, comprising:

enclosing a substrate in a semi-porous microcapsule comprising a hydrogel shell and an aqueous core;
washing the semi-porous microcapsule in a first reaction buffer comprising first reaction reagents such that the first reaction buffer substantially replaces any prior microcapsule buffer and such that first reaction reagents enter the microcapsule aqueous core;
performing the first reaction on the substrate to form a first substrate product in the semi-porous microcapsule;
washing the semi-porous microcapsule in a second reaction buffer comprising second reaction reagents such that the second reaction buffer substantially replaces the first reaction buffer at the microcapsule aqueous core and such that second reaction reagents replace the first reaction reagents in the microcapsule aqueous core;
performing the second reaction on the first substrate product in the semi-porous microcapsule aqueous core to form a second substrate product.

2 more elements

Dependencies are inferred from the claim text.

Systems and methods for encapsulation and multi-step processing of biological samples

Life of this patent

Systems and methods for encapsulation and multi-step processing of biological samples

Life of this patent

Abstract

Claims30