TEMPLATE PARTICLES WITH MICROPORES AND NANOPORES

1 . A composition comprising:

a plurality of hydrogel particles suspended in an aqueous liquid, each hydrogel particle comprising a mesh of cross-linked polymers, wherein each hydrogel particle comprises:

(i) a plurality of micropores extending through the mesh of cross-linked polymers, each micropore having an open interior volume having a dimension of at least about a micron; and/or

(ii) a nanoporous structure in the mesh of cross-linked polymers, wherein the hydrogel mesh has a mesh size of at least about 200 nm, wherein the nanoporous structure comprises at least one open interior volume in the hydrogel mesh;

wherein the particle is loaded with a reaction reagent; and

wherein the aqueous liquid permeates the interior volume of the micropores and/or nanoporous structure allowing analytes in the fluid to access the reagent.

2 . The composition of claim 1 , wherein each particle comprises a plurality of micropores.

3 . The composition of claim 2 , wherein each particle further comprises a nanoporous structure.

4 . The composition of claim 1 , wherein each particle comprises a nanoporous structure.

5 . The composition of claim 1 , wherein the reagents comprise one or more of enzymes, enzyme cofactors, nucleotides, polynucleotides, amino acids, peptides, proteins, probes, primers, salts, ions, buffers, labels, dyes, antibodies, polymers, and carbohydrates.

6 . The composition of claim 5 , wherein the reagents comprise one or more target capture moiety.

7 . The composition of claim 6 , wherein the target capture moiety captures one or more of circulating cells, cellular components, cell-free nucleic acids, extracellular vesicles, protein antigens, prokaryotic cells, fungi, viruses, and combinations thereof.

8 . The composition of claim 5 , wherein the reagents comprise reagents for one or more of nucleic acid synthesis, transcription, reverse transcription, and cell lysis.

9 . The composition of claim 5 , wherein one or more of the reagents are covalently linked to the particle.

10 . A method for performing a bioassay, the method comprising:

combining template particles with samples in a first fluid, wherein the template particles are hydrogel particles suspended in the first fluid, each hydrogel particle comprising a mesh of cross-linked polymers, wherein each hydrogel particle is loaded with a reaction reagent and comprises:

(i) a plurality of micropores extending through the mesh of cross-linked polymers, each micropore having an open interior volume having a dimension of at least about a micron; and/or

(ii) a nanoporous structure in the mesh of cross-linked polymers, wherein the hydrogel mesh has a mesh size of at least 200 nm in length, wherein the nanoporous structure comprises at least one open interior volume in the hydrogel mesh;

adding a second fluid immiscible to the first fluid; and

shearing the fluids to generate a plurality of monodispersed droplets simultaneously that contain a single one of the template particles and one or more of the samples.

11 . The method of claim 10 , wherein the first liquid permeates the interior volume of the micropores and/or nanoporous structure allowing analytes in the first fluid to access the reaction reagent.

12 . The method of claim 11 , wherein in the monodisperse droplets, the interior volume is occupied by at least the first fluid and one or more of the reagents.

13 . The method of claim 10 , wherein the samples comprise at least one of circulating cells, cellular components, cell-free nucleic acids, extracellular vesicles, protein antigens, prokaryotic cells, fungi, viruses, and combinations thereof.

14 . The method of claim 13 , wherein the samples are cells and the sample in each of the plurality of monodisperse droplets is a single cell.

15 . A method for producing cross-linked template particles, the method comprising:

preparing an aqueous phase fluid comprising an acrylamide/bisacrylamide copolymer matrix, wherein the aqueous phase fluid comprises at least about 3.5 wt % acrylamide/bisacrylamide; and

co-flowing the aqueous phase fluid and a fluid immiscible to the aqueous phase fluid through a droplet generation device,

wherein the resulting template particles have an effective hydrogel mesh size less than 200 nm in length.

17 . The method of claim 16 , wherein the effective hydrogel mesh size is modulated via a ratio of acrylamide monomers to bisacrylamide monomers in the copolymer matrix.

18 . A method for producing cross-linked template particles, the method comprising:

preparing an aqueous phase fluid comprising acrylamide monomers and PEG; and

co-flowing the aqueous phase fluid and a fluid immiscible to the aqueous phase fluid through a droplet generation device,

wherein the resulting template particles comprise micropores having an open interior volume having a dimension of at least about a micron.

19 . The method of claim 18 , wherein the aqueous phase fluid comprises at least about 4 wt % and 2 wt % PEG.

20 . The method of claim 18 , further comprising washing the PEG from the template particles.