US 11,860,076 B2
Systems and methods for encapsulation and multi-step processing of biological samples
Linas Mazutis, Vilnius (LT); Greta Stonyte, Vilnius (LT); Karolis Leonavicius, Vilnius (LT); and Ausra Zelvyte, Vilnius (LT)
Assigned to Vilnius University, Vilnius (LT); and Droplet Genomics, Vilnius (LT)
Filed by Vilnius University, Vilnius (LT); and Droplet Genomics, Vilnius (LT)
Filed on Jul. 21, 2020, as Appl. No. 16/934,045.
Claims priority of provisional application 62/863,881, filed on Jun. 20, 2019.
Prior Publication US 2020/0400538 A1, Dec. 24, 2020
Int. Cl. G01N 1/40 (2006.01); C12Q 1/68 (2018.01); B01J 13/04 (2006.01); B01J 13/20 (2006.01); C12Q 1/6806 (2018.01)
CPC G01N 1/4077 (2013.01) [B01J 13/046 (2013.01); B01J 13/206 (2013.01); C12Q 1/6806 (2013.01)] 15 Claims
 
1. A method for isolating species in microscopic compartments comprising a semi-permeable shell and an aqueous inner phase, and processing encapsulated species in a series of reactions/procedures, wherein the method comprises:
forming a liquid droplet on a microfluidic chip, wherein a first aqueous phase, a second aqueous phase, and a carrier oil are injected into the chip to form the liquid droplet, and wherein the species is in the first or second aqueous phase;
forming an aqueous two-phase system passively, wherein the first and second aqueous phases are miscible and form an inner phase and an outer phase, respectively;
hardening the outer phase using physical or chemical stimulus, thereby creating a hardened shell encapsulating the species in the aqueous inner phase; and
performing one or more biological or biochemical reactions on the encapsulated species in microscopic compartments, wherein performing the one or more biological or biochemical reactions on the encapsulated species in the microscopic compartments comprises resuspending the microscopic compartments in assay reagents, and wherein the hardened shell provides a semi-permeable shell allowing for exchange of compounds having molecular weights of less than 200,000 MW through the shell and retention of compounds having molecular weight greater than 300,000 MW.