Device and method for analyzing a biological sample

Abstract

Systems for analyzing biological components are provided. The systems can include a fluidic device and an energy source in communication with the fluidic device. The energy source can supply energy to the fluidic device to form a polymer matrix on or adjacent to a biological component within the fluidic device. Methods of analyzing biological components using the systems are also provided.

Description

[0001] Cross-references This application claims the benefit of U.S. Provisional Application No. 63 / 609,847, filed December 13, 2023, which is incorporated herein by reference in its entirety. Background Technology

[0002] In the field of cell biology, single-cell analysis can encompass the study of genomics, transcriptomics, proteomics, metabolomics, and cell-cell interactions at the single-cell level. Due to the heterogeneity present in both eukaryotic and prokaryotic cell populations, analyzing single cells makes it possible to uncover mechanisms that are undetectable when studying large cell populations. In single-cell analysis, changes in a single cell can be tracked or observed at the level of genes, proteins, or other cellular components. For example, in cancer, cells may mutate, and understanding changes at the genetic level in cancer can be meaningful. Single-cell sequencing can be used to observe patterns of somatic mutations and copy number aberrations. Summary of the Invention

[0003] This paper recognizes the need to compartmentalize biological samples to perform one or more assays on individual components within each compartment. One or more assays can be performed with or without additional processing of individual components (e.g., without nucleotide amplification steps), while preserving the spatial information of each component. Compartments can be generated or deconstructed as needed to locate or release targeted components of the biological sample.

[0004] In some embodiments, this document provides a system comprising a jetting device containing one or more biological components and one or more polymer precursors. The system may also include at least one energy source in communication with the jetting device. In some embodiments, the at least one energy source supplies energy to the jetting device to cause the one or more polymer precursors to form at least one polymer matrix on or adjacent to the biological component.

[0005] In some embodiments, the polymer matrix forms a plurality of adjacent gel structures, wherein said gel structures are degradable. In a further embodiment, degradation of at least a portion of at least two adjacent gel structures allows for the merging of the contents of said at least two adjacent gel structures.

[0006] Another aspect of this disclosure provides a non-transitory computer-readable medium comprising machine-executable code that, when executed by one or more computer processors, implements any of the methods described above or elsewhere herein.

[0007] Another aspect of this disclosure provides a system comprising one or more computer processors and computer memory coupled thereto. The computer memory includes machine-executable code that, when executed by the one or more computer processors, implements any of the methods described above or elsewhere herein.

[0008] Further aspects and advantages of this disclosure will become readily apparent to those skilled in the art from the following detailed description, in which only illustrative embodiments of this disclosure are shown and described. As will be understood, this disclosure is capable of other and different embodiments, and certain details thereof can be modified in various obvious respects without departing from this disclosure. Therefore, the drawings and description should be considered illustrative rather than restrictive.

[0009] Provided in some embodiments, this document describes a method for processing chemical or biological materials, comprising: (a) inputting (i) the chemical or biological material and (ii) one or more polymer precursors into a jetting device; (b) supplying a stimulus to the jetting device to generate one or more polymer matrices, wherein the one or more polymer matrices comprise: (i) a first gel structure from the one or more polymer precursors within the jetting device, wherein the first gel structure at least partially encapsulates the chemical or biological material, and wherein the first gel structure comprises a first internal region; and (ii) a second gel structure adjacent to the first gel structure, wherein the second gel structure comprises a second internal region; and (c) degrading the first gel structure, the second gel structure, or a portion thereof to generate a third internal region, wherein the third internal region comprises the first internal region of the first gel structure and the second internal region of the second gel structure.

[0010] In some embodiments, the first gel structure and the second gel structure each comprise one or more polymer matrix walls. In some embodiments, the first gel structure and the second gel structure share the polymer matrix wall of the one or more polymer matrix walls. In some embodiments, a portion of the first gel structure, the second gel structure, or both degraded in (c) includes the shared polymer matrix wall.

[0011] In one aspect, this document describes a method for processing chemical or biological materials, comprising: (a) introducing (i) the chemical or biological material and (ii) one or more polymeric precursors into a jetting device; (b) supplying a stimulus to the jetting device to generate one or more polymeric matrices from the one or more polymeric precursors within the jetting device, wherein the polymeric matrix of the one or more polymeric matrices at least partially encapsulates the chemical or biological material, and wherein the polymeric matrix is ​​formed within less than 3 seconds after the one or more polymeric precursors come into contact with the stimulus.

[0012] In some embodiments, the polymer matrix is ​​formed within less than 2 seconds after the one or more polymer precursors are contacted with the energy. In some embodiments, the polymer matrix is ​​formed within less than 1 second after the one or more polymer precursors are contacted with the energy. In some embodiments, the polymer matrix is ​​formed within less than 0.5 seconds after the one or more polymer precursors are contacted with the energy.

[0013] In one aspect, this document describes a method for treating chemical or biological materials, comprising (a) introducing (i) the chemical or biological material and (ii) one or more polymeric precursors into a jetting device; (b) supplying a stimulus to the jetting device to generate one or more polymeric matrices from the one or more polymeric precursors within the jetting device, wherein the polymeric matrix of the one or more polymeric matrices at least partially encapsulates the chemical or biological material; and (c) performing one or more operations on the chemical or biological material within the polymeric matrix, wherein the one or more operations include a temporal duration of at least one day, during which no more than 20% of the polymeric matrix degrades.

[0014] In some embodiments, no more than 15% of the polymer matrix degrades during the time interval. In some embodiments, no more than 10% of the polymer matrix degrades during the time interval. In some embodiments, no more than 5% of the polymer matrix degrades during the time interval. In some embodiments, no more than 1% of the polymer matrix degrades during the time interval. In some embodiments, the time interval includes at least three days. In some embodiments, the time interval includes at least five days. In some embodiments, the time interval includes at least seven days. In some embodiments, the time interval includes at least fourteen days.

[0015] In one aspect, this document describes a method for processing chemical or biological materials, comprising: (a) introducing (i) one or more chemical or biological materials and (ii) one or more polymeric precursors into a jetting device; (b) supplying a stimulus to the jetting device to generate one or more polymeric matrices from the one or more polymeric precursors within the jetting device, wherein the one or more polymeric matrices at least partially encapsulate the one or more chemical or biological materials; and (c) performing one or more operations on the one or more chemical or biological materials within the one or more polymeric matrices, wherein the one or more operations comprise a time interval of at least one day, during which at least 80% of the one or more chemical or biological materials are viable. In some embodiments, at least 85% of the one or more chemical or biological materials are viable during the time interval. In some embodiments, at least 90% of the one or more chemical or biological materials are viable during the time interval. In some embodiments, at least 95% of the one or more chemical or biological materials are viable during the time interval. In some embodiments, at least 99% of the one or more chemical or biological materials are viable during the time interval. In some embodiments, the time interval comprises at least three days. In some embodiments, the time interval includes at least five days. In some embodiments, the time interval includes at least seven days. In some embodiments, the time interval includes at least fourteen days.

[0016] In one aspect, this document describes a method for processing chemical or biological materials, comprising: (a) feeding (i) one or more chemical or biological materials and (ii) one or more polymer precursors into a jet device; and (b) maintaining jet communication between the one or more chemical or biological materials and the polymer precursors for a duration of at least ten minutes, wherein at least 80% of the one or more chemical or biological materials are viable during the duration.

[0017] In some embodiments, at least 85% of the one or more chemical or biological materials are active during the time interval. In some embodiments, at least 90% of the one or more chemical or biological materials are active during the time interval. In some embodiments, at least 95% of the one or more chemical or biological materials are active during the time interval. In some embodiments, at least 99% of the one or more chemical or biological materials are active during the time interval. In some embodiments, the time interval includes at least 30 minutes. In some embodiments, the time interval includes at least one hour. In some embodiments, the time interval includes at least two hours. In some embodiments, the time interval includes at least three hours. In some embodiments, the time interval includes at least one day. In some embodiments, the method further includes supplying a stimulus to the jetting device to generate one or more polymer matrices from the one or more polymer precursors within the jetting device, wherein the one or more polymer matrices at least partially encapsulate the one or more chemical or biological materials. In some embodiments, the stimulus includes light. In some embodiments, the one or more polymer matrices comprise a polymeric form of a cPEG monomer.

[0018] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising the structure of structure (I): .

[0019] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (II): (II).

[0020] In some embodiments, n is between about 0 and about 10. In some embodiments, the one or more polymer precursors comprise at least 40 µL of solution.

[0021] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (III): (III).

[0022] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (IV): .

[0023] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising a structure (V): .

[0024] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (VI): .

[0025] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (VII): (VII)

[0026] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (VIII): .

[0027] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising a structure (IX): (IX).

[0028] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (X): (X).

[0029] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (XI): .

[0030] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (XII): .

[0031] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (XIII): .

[0032] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising: an oligomeric domain having three or more arms, wherein each arm of the oligomeric domain comprises a degradable functional group and a crosslinkable functional group, wherein the crosslinkable functional group of one of the three or more arms is configured to crosslink with another crosslinkable functional group of another polymer precursor in response to a first stimulus to obtain the polymeric form of the monomer, and wherein the degradable functional group is configured to dissolve the polymeric form of the monomer in response to a second stimulus being cleaved.

[0033] In some implementations, the oligomer domain comprises four or more arms.

[0034] In some embodiments, the one or more polymer matrices comprise degradable functional groups. In some embodiments, the degradable functional groups include disulfides, β-thioether esters, amide hydroxymethyl and vicinal diols, vicinal diols, alginate backbones, dextran backbones, chitosan backbones, hyaluronic acid backbones, chondroitin sulfate backbones, or carboxymethyl cellulose backbones, or combinations thereof. In some embodiments, the one or more polymer matrices comprise hydrogels. In some embodiments, the hydrogels comprise cPEG, cSEL-BTEEC, cSEL-DHEBA, cSEL-diol, cSEL-alginate, cSEL-dextran, cSEL-chitosan, cSEL-hyaluronic acid, cSEL-chondroitin sulfate, or cSEL-cellulose, or combinations thereof. In some embodiments, the one or more polymer matrices comprise degrading agents. In some embodiments, the degradation reagent includes DTT, TCEP, BME, GSH, DMEM, RPMI, PBS buffer, DMEM, RPMI, PBS buffer, sodium (meta)periodate, alginate lyase (enzyme), dextranase, lysozyme and chitinase, hyaluronidase, chondroitinase or cellulase or combinations thereof.

[0035] In some embodiments, the one or more polymer matrices comprise at least one β-thioether ester. In some embodiments, the one or more polymer matrices comprise a PEG moiety containing a β-thioether ester. In some embodiments, the β-thioether ester is formed by reacting an acrylate with a thiol. In some embodiments, the one or more polymer matrices are degraded overnight in the presence of a commonly used cell culture medium, thereby providing a biocompatible degradation pathway for selective retention experiments. In some embodiments, the one or more polymer matrices comprise a Michael donor. In some embodiments, the Michael donor is a PEG-thiol. In some embodiments, the one or more polymer matrices comprise a cSEL β-thioether ester having one β-thioether ester per arm. In some embodiments, the one or more polymer matrices comprise any material formed of PEG containing a Michael acceptor chain. In some embodiments, the Michael acceptor chain comprises PEG-acrylamide, PEG-vinyl sulfone, PEG-maleimide, or PEG-carbonylacrylic acid, or any combination thereof.

[0036] In some embodiments, the one or more polymer matrices are degraded by cleavage of disulfide bonds. In some embodiments, the disulfide bonds are cleaved by one or more reducing agents. In some embodiments, the one or more reducing agents include DTT, TCEP, BME, or GSH, or any combination thereof. In some embodiments, the one or more polymer matrices comprise one or more arms, each containing one or more amides. In some embodiments, the one or more polymer matrices are degraded by oxidative cleavage of vicinal diols with (met)periodate. In some embodiments, the method further comprises: (a) using a caged workflow; (b) performing selective cell retention when used with cPEG or other hydrogels having an orthogonal degradation mechanism; or (c) performing long-term cell incubation using a reducing agent that does not affect cage stability in the cell culture medium. In some embodiments, the one or more polymer matrices comprise vicinal diol functionality that can be cleaved via oxidation of one or more hydroxyl groups. In some embodiments, the one or more polymer matrices are used to formulate stable hydrogels. In some embodiments, the one or more polymer matrices comprise a polymeric form of a photocleavable 4-arm PEG monomer. In some embodiments, the one or more polymer matrices are photodegraded via an o-nitrobenzyl moiety. In some embodiments, the one or more polymer matrices comprise a polymeric form of a coumarin-based photodegradable monomer. In some embodiments, the one or more polymer matrices comprise a 4-arm PEG-acrylamide containing one or more disulfides. In some embodiments, the one or more polymer matrices contain one or more cage disulfide bonds in the formation of hydrogel cages, wherein the hydrogel cages are degraded using light and a photoinitiator. In some embodiments, the one or more polymer matrices enable hydrogel formation, and wherein the hydrogel enables spatiotemporal control of hydrogel cage degradation, thus enabling selective retention of cells with a single hydrogel formulation. In some embodiments, the one or more polymer matrices enable hydrogel formation, and wherein the hydrogel, using light and a photoinitiator as a reagent, enables the hydrogel to selectively degrade cages and release their contents.

[0037] In some embodiments, the one or more polymer precursors enable the formation of hydrogels up to 2 mm thick, and the hydrogels can be formed within 120 s with low light intensity (e.g., 10 mW / cm² at 265 nm). 2 Approximately 50 to 1000 mW / cm 2 Or approximately 100 to 700 mW / cm 2 ) is degraded.

[0038] In some implementations, when exposed to light, photogenerated free radicals initiate multiple fracture and disulfide exchange reactions, thereby allowing and promoting photodeformation, photowelding, and photodegradation of the one or more polymer matrices.

[0039] In some embodiments, the one or more polymer precursors enable the formation of hydrogels, wherein the hydrogels exhibit chemical or physical changes in response to external stimuli.

[0040] In some embodiments, the one or more polymer matrices include photoinstantaneous nitrobenzyl esters that cleave upon photon absorption, thereby allowing the user to exogenously control the degradation of the one or more polymer matrices.

[0041] In some embodiments, the one or more polymer precursors enable hydrogel formation, and the method further includes controlling the network degradation of the hydrogel by means of the concentration of a photoinitiator infused into the hydrogel.

[0042] In some embodiments, the one or more polymer precursors comprise thiol groups, which are quenched after the formation of the one or more polymer matrices.

[0043] In some implementations, higher concentrations of bisacrylamide in one or more polymer matrices result in larger pore sizes.

[0044] In some embodiments, the degradation of the one or more polymer matrices depends on the type and number of encapsulated cells, or the fraction of disulfide moieties present in the one or more polymer matrices. In some embodiments, the grafting efficiency of the one or more polymer matrices is at least 96%.

[0045] In some embodiments, one or more polymer precursors are in a solution with a pH of at least 8.0.

[0046] In some embodiments, the one or more polymer precursors include a porogen, and increasing the porogen loading results in an increased pore size in the one or more polymer matrices. In some embodiments, the increased porogen concentration increases the diffusion rate by about 20%. In some embodiments, the increased pore size increases the diffusion rate of the fluorescently labeled antibody into the one or more polymer matrices. In some embodiments, the fluorescently labeled antibody is CD56 PE. In some embodiments, the one or more polymer precursors are photopolymerized via photocrosslinking by UV light.

[0047] Another aspect of this disclosure provides a flow pool for sequencing genes, comprising: a plurality of cells, a plurality of nanopores, and one or more polymer precursors; wherein a polymer matrix is ​​formed by the one or more polymer precursors within less than 10 seconds after the polymer precursors are contacted with energy.

[0048] In some embodiments, the method includes the chemical or biological material comprising a first cell in the first internal region and a second cell in the second internal region.

[0049] In some embodiments, the method includes the first cell comprising one of killer cells, activating cells, and inhibitory cells, and wherein the first cell interacts with the second cell after the degradation in (c).

[0050] In some embodiments, the method includes the chemical or biological material comprising polymer beads containing one or more oligonucleotides disposed thereon.

[0051] In some embodiments, the method includes the chemical or biological material comprising one or more cells or components thereof. In some embodiments, the method further includes one or more polymer precursors comprising: (a) a macromonomer, (b) a photoinitiator, and (c) a porogen. In some embodiments, the method further includes the degradation in (c) caused by cleavage of one or more disulfide bonds in the first gel structure, the second gel structure, or both.

[0052] Another aspect of this disclosure provides a polymer precursor comprising: an oligomeric domain having three or more arms, wherein each arm of the oligomeric domain comprises a degradable functional group and a crosslinkable functional group, wherein the crosslinkable functional group of one of the three or more arms is configured to crosslink with another crosslinkable functional group of another polymer precursor in response to a first stimulus to obtain a polymeric form of the polymer precursor, and wherein the degradable functional group is configured to dissolve the polymeric form of the polymer precursor in response to a second stimulus being cleaved.

[0053] In some embodiments, the oligomeric domain comprises four or more arms. In some embodiments, the oligomeric domain is hydrophilic. In some embodiments, the oligomeric domain comprises poly(ethylene glycol), poly(N-vinylpyrrolidone), poly(acrylic acid), poly(methacrylic acid), poly(vinyl alcohol), poly(L-lysine), poly(2-ethyl-2-oxazoline), poly(maleic acid), poly(vinyl phosphate), poly(acrylamide), poly(vinylamine), poly(ethylene oxide-co-propylene oxide), poly(N-isopropylacrylamide), poly(vinylphosphonic acid), poly(2-vinyl-1-methylpyridinium bromide), poly(N,N-diethylacrylamide), poly(N,N-dimethylacrylamide), poly(2-hydroxyethyl acrylate), poly(2-hydroxy methacrylate-co-methacrylic acid), poly(propylene oxide), poly(2-(dimethylamino)ethyl methacrylate), or poly(N-(2-hydroxypropyl)methacrylamide) or combinations thereof. In some embodiments, the oligomer domain comprises poly(ethylene glycol). In some embodiments, the oligomer domain comprises poly(N-vinylpyrrolidone). In some embodiments, the oligomer domain comprises poly(acrylic acid). In some embodiments, the oligomer domain comprises poly(methacrylic acid). In some embodiments, the oligomer domain comprises poly(vinyl alcohol). In some embodiments, the oligomer domain comprises poly(L-lysine). In some embodiments, the oligomer domain comprises poly(2-ethyl-2-oxazoline). In some embodiments, the oligomer domain comprises poly(maleic acid). In some embodiments, the oligomer domain comprises poly(vinylphosphonic acid). In some embodiments, the oligomer domain comprises poly(acrylamide). In some embodiments, the oligomer domain comprises poly(vinylamine). In some embodiments, the oligomer domain comprises poly(ethylene oxide-co-propylene oxide).

[0054] In some embodiments, the oligomeric domain comprises poly(N-isopropylacrylamide). In some embodiments, the oligomeric domain comprises poly(vinylphosphonic acid). In some embodiments, the oligomeric domain comprises poly(2-vinyl-1-methylpyridinium bromide). In some embodiments, the oligomeric domain comprises poly(N,N-diethylacrylamide). In some embodiments, the oligomeric domain comprises poly(N,N-dimethylacrylamide). In some embodiments, the oligomeric domain comprises poly(2-hydroxyethyl acrylate). In some embodiments, the oligomeric domain comprises poly(2-hydroxy methacrylate-co-methacrylic acid). In some embodiments, the oligomeric domain comprises poly(propylene oxide). In some embodiments, the oligomeric domain comprises poly(2-(dimethylamino)ethyl methacrylate). In some embodiments, the oligomeric domain comprises poly(N-(2-hydroxypropyl)methacrylamide).

[0055] In some embodiments, the degradable functional group includes disulfides, β-thioether esters, amide hydroxymethyl groups, o-diols, peptides, polysaccharides (e.g., alginate, dextran, chitosan, chondroitin), oligosaccharides, o-nitrobenzyl groups, coumarins, acetals, ketals, esters, orthoesters, acid anhydrides, imines, hydrazones, carbonates, carbamates, azo dyes, phosphates, cinnamoyl groups, or benzoyl ethers, or combinations thereof. In some embodiments, the degradable functional group includes disulfides. In some embodiments, the degradable functional group includes β-thioether esters. In some embodiments, the degradable functional group includes disulfides, β-thioether esters, amide hydroxymethyl groups, vicinal diols, peptides, polysaccharides (e.g., alginate, dextran, chitosan, chondroitin), vicinal nitrobenzyl groups, coumarin, acetals, ketals, esters, orthoesters, acid anhydrides, imines, hydrazones, carbonates, phosphates, cinnamoyl groups, or benzoyl ethers, or combinations thereof. In some embodiments, the degradable functional group includes disulfides. In some embodiments, the degradable functional group includes β-thioether esters. In some embodiments, the degradable functional group includes disaccharides. In some embodiments, the degradable functional group includes amide hydroxymethyl groups. In some embodiments, the degradable functional group includes vicinal diols. In some embodiments, the degradable functional group includes peptides. In some embodiments, the degradable functional group includes one or more polysaccharides.

[0056] In some embodiments, the one or more polysaccharides comprise alginate. In some embodiments, the one or more polysaccharides comprise dextran. In some embodiments, the one or more polysaccharides comprise chitosan. In some embodiments, the one or more polysaccharides comprise chondroitin. In some embodiments, the degradable functional group comprises o-nitrobenzyl. In some embodiments, the degradable functional group comprises coumarin. In some embodiments, the degradable functional group comprises acetal. In some embodiments, the degradable functional group comprises ketal. In some embodiments, the degradable functional group comprises ester. In some embodiments, the degradable functional group comprises orthoester. In some embodiments, the degradable functional group comprises anhydride. In some embodiments, the degradable functional group comprises imine. In some embodiments, the degradable functional group comprises hydrazone.

[0057] In some embodiments, the degradable functional group comprises a carbonate. In some embodiments, the degradable functional group comprises a phosphate ester. In some embodiments, the degradable functional group comprises a cinnamoyl group. In some embodiments, the degradable functional group comprises benzoin. In some embodiments, the degradable functional group comprises an azo group. In some embodiments, the degradable functional group comprises a carbamate. In some embodiments, the degradable functional group comprises an oligosaccharide. In some embodiments, the oligosaccharide comprises 2, 3, or 4 monomer units (e.g., 2, 3, or 4 hexoses, pentoses, heptoses, combinations thereof, etc.). In some embodiments, the oligosaccharide is a disaccharide.

[0058] In some embodiments, the crosslinkable functional group includes acrylates, methacrylates, acrylamides, methacrylamides, norbornel, styrene, vinyl ethers, vinylpyrrolidones, vinyl esters, maleimides, allyl groups, alkynes, azides, thiols, olefins, or epoxides or combinations thereof.

[0059] In some embodiments, the crosslinkable functional group comprises acrylate. In some embodiments, the crosslinkable functional group comprises methacrylate. In some embodiments, the crosslinkable functional group comprises acrylamide. In some embodiments, the crosslinkable functional group comprises methacrylamide. In some embodiments, the crosslinkable functional group comprises norbornyl. In some embodiments, the crosslinkable functional group comprises styrene. In some embodiments, the crosslinkable functional group comprises vinyl ether. In some embodiments, the crosslinkable functional group comprises vinylpyrrolidone.

[0060] In some embodiments, the crosslinkable functional group comprises a vinyl ester. In some embodiments, the crosslinkable functional group comprises a maleimide. In some embodiments, the crosslinkable functional group comprises an allyl group. In some embodiments, the crosslinkable functional group comprises an alkyne. In some embodiments, the crosslinkable functional group comprises an azide. In some embodiments, the crosslinkable functional group comprises a thiol. In some embodiments, the crosslinkable functional group comprises an olefin. In some embodiments, the crosslinkable functional group comprises an epoxide. In some embodiments, the first stimulus comprises an energy source. In some embodiments, the energy source comprises a light source. In some embodiments, the wavelength of the light source is between 300 and 500 nm. However, the wavelength can be selected to suit a particular photoinitiator. For example, a wavelength of about 275 nm can be selected for 2-hydroxy-2-methyl-phenyl-propane-1-one; a wavelength of about 405 nm can be selected for lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP); a wavelength of about 635 nm can be selected for 1,4-bis(pentylamino)anthraquinone; and so on.

[0061] In some embodiments, the second stimulus comprises an energy source in the presence of a photoinitiator and in the absence of a polymer precursor. In some embodiments, the energy source comprises a light source. In some embodiments, the wavelength of the light source is between 300 and 500 nanometers (nm). In some embodiments, the wavelength of the light source is between 380 nm and 420 nm. In some embodiments, the wavelength of the light source is between about 250 and 700 nm, about 250 and 300 nm, about 250 and 400 nm, about 300 and 450 nm, or about 400 and 700 nm.

[0062] In some embodiments, the second stimulus includes the introduction of a degradation agent. In some embodiments, the degradation agent includes dithiothreitol (DTT).

[0063] In some embodiments, the degradation agent comprises tris(2-carboxyethyl)phosphine (TCEP). In some embodiments, the degradation agent comprises 2-mercaptoethanol (BME). In some embodiments, the degradation agent comprises glutathione reductase (GSH). In some embodiments, the degradation agent comprises DMEM. In some embodiments, the degradation agent comprises RPMI. In some embodiments, the degradation agent comprises PBS buffer. In some embodiments, the degradation agent comprises sodium (meta)periodate. In some embodiments, the degradation agent comprises alginate lyase. In some embodiments, the degradation agent comprises dextranase. In some embodiments, the degradation agent comprises lysozyme. In some embodiments, the degradation agent comprises chitinase. In some embodiments, the degradation agent comprises hyaluronidase. In some embodiments, the degradation agent comprises chondroitinase.

[0064] In some embodiments, the degradation agent includes cellulase.

[0065] Another aspect of this disclosure provides a kit comprising: (a) a porogen; and (b) a polymer precursor described anywhere in this disclosure.

[0066] In some embodiments, the porogen includes polyethylene glycol.

[0067] In some embodiments, the ratio of the porogen to the polymer precursor by weight is between about 1:1 and 1:6. In some embodiments, the ratio of the porogen to the polymer precursor by weight is between about 1:2 and 1:5. In some embodiments, the ratio of the porogen to the polymer precursor by weight is between about 1:3 and 1:4. In some embodiments, the ratio of the porogen to the polymer precursor by weight is about 1:3.5.

[0068] In some embodiments, (a) the porogen and (b) the polymer precursor are contained in a single container. In some embodiments, the kit further comprises (c) a photoinitiator contained in another container.

[0069] In some embodiments, the photoinitiator includes one of lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), Irgacure 2959, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide (TPO) nanoparticles, 2,2'-azobis[2-methyl-N-(2-hydroxyethyl)promionamide] (VA-086), BAPO-Oli, BAPO-Ona, eosin Y, riboflavin, and combinations thereof.

[0070] In some embodiments, the oligomeric domain further comprises an amide moiety. In some embodiments, the oligomeric domain further comprises a β-thioether moiety.

[0071] In one aspect, this document describes a method for treating chemical or biological materials, comprising: (a) introducing (i) the chemical or biological material and (ii) one or more polymer precursors into a jetting device, wherein the one or more polymer precursors comprise a porogen, a macromonomer, and a photoinitiator; (b) supplying an optical stimulus to the jetting device to generate two or more polymer matrices, wherein the two or more polymer matrices comprise: (i) a first hydrogel structure formed within the jetting device, wherein the first hydrogel structure encapsulates the chemical or biological material and wherein the first hydrogel structure includes a first internal region; and (ii) a second hydrogel structure formed within the jetting device, wherein the second hydrogel structure encapsulates the chemical or biological material and wherein the first hydrogel structure includes a second internal region; and (c) introducing a solution containing the photoinitiator into the jetting device; and (d) supplying the optical stimulus to the jetting device to selectively degrade at least one of the first hydrogel structure or the second hydrogel structure.

[0072] In some embodiments, feeding the solution containing the photoinitiator into the jet apparatus includes removing the porogen and the macromonomer from the jet apparatus. In some embodiments, the macromonomer comprises a disulfide moiety, wherein two or more polymer matrices contain the disulfide moiety, the disulfide moiety being configured to be cleaved upon exposure to the supplied optical stimulus.

[0073] In some embodiments, the at least partial encapsulation of the chemical or biological material is a complete encapsulation of the chemical or biological material. In some embodiments, the gel structure is a hydrogel structure. In some embodiments, the viable one or more biological materials include living cells.

[0074] Incorporation All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference as if each individual publication, patent, or patent application were expressly and individually indicated to be incorporated by reference. Where a publication, patent, or patent application incorporated by reference contradicts the disclosure contained in this specification, this specification is intended to supersede and / or give precedence to any such contradictory material. Attached Figure Description

[0075] The novel features of the invention are particularly set forth in the appended claims. A better understanding of the features and advantages of the invention will be obtained by referring to the following detailed description and accompanying drawings (also referred to herein as “Figure” and “FIG.”) illustrating illustrative embodiments utilizing the principles of the invention, in which: Figures 1A-1E Illustrative examples of polymer matrix compartment formation and encapsulation of one or more biological components in a flow cell over a certain period of time, according to some embodiments, are shown. Figure 2A The effect of buffer concentration in the polymer matrix precursor solution on the degradation rate of the polymer matrix is ​​shown according to some embodiments; Figure 2B The following embodiments are shown: the polymer matrix is ​​cleaved by a degradation agent in buffers of different pH values. Figure 3 The following embodiments are shown: the polymer matrix is ​​cleaved by a degradation agent in buffers of different pH values. Figure 4A It is an illustrative graph comparing the gel lysis time in minutes on the Y-axis with the pH of the working solution on the X-axis, based on some implementation schemes; Figure 4B It is an illustrative graph comparing the gel lysis time in minutes on the Y-axis with the pH of the working solution on the X-axis, based on some implementation schemes; Figures 5A-5D Illustrative examples of micrographs showing the encapsulation of a polymer matrix compartment and one or more biological components in a flow cell over a period of time after formation / curing, according to some embodiments; Figure 5E Illustrative examples of fluorescence micrographs of polymer matrix compartments encapsulated with one or more biological components in a flow cell, 14 days after formation / curing, are shown according to some embodiments. Figures 6A-6C Illustrative examples of micrographs showing the encapsulation of a polymer matrix compartment and one or more biological components in a flow cell some time after self-cageing, according to some embodiments; Figures 6D-6F Illustrative examples of micrographs of polymer matrix compartments and encapsulation of one or more biological components in a flow cell after cage formation for a period of time, according to some embodiments, wherein the biological components are exposed to the polymer precursor for 3 hours prior to cage formation. Figure 7This is an illustrative graph comparing the % background intensity on the Y-axis with the diffusion time (min) of CD56 PE (dye-labeled antibody) across the polymer matrix on the X-axis, based on some implementation schemes. Figures 8A-8D Illustrative examples of micrographs showing polymer-matrix compartment encapsulation of one or more biological components in a flow cell according to some embodiments; Figures 9A-9B Illustrative examples of precursor solution compositions by volume for forming polymer matrix compartments configured to encapsulate one or more biological components in a flow cell, according to some embodiments, are shown. Figure 10A This is an illustrative graph comparing the % background intensity on the Y-axis with the diffusion time (min) of CD56 PE to the polymer matrix on the X-axis, based on some implementation schemes. Figure 10B It is an illustrative graph that compares the % background intensity on the Y-axis at fixed time intervals with the concentration of the polymer precursor stock solution on the X-axis, according to some implementation schemes. Figure 11 Illustrative examples of precursor solution compositions by volume for forming polymer matrix compartments configured to encapsulate one or more biological components in a flow cell, according to some embodiments, are shown. Figures 12A-12D Examples of polymer matrices during cage formation, before degradation, after degradation, and after rinsing are shown according to some embodiments; Figures 12E-12G An example is shown where, according to some embodiments, a portion of the polymer matrix cage is degraded to incorporate elements in the polymer matrix; Figure 13 Computer systems, programmed or otherwise configured to implement the methods provided herein, are shown according to some embodiments; Figure 14A The diagram shows a portion of a system, such as those presented herein, including an energy source, according to some implementation schemes. Figure 14B The diagram shows a polymer matrix formed around a biological component in a part of a system as provided herein, according to some embodiments.

[0076] Figure 14C This paper illustrates a method for forming a polymer matrix around a biological component in a system as provided herein, according to some embodiments.

[0077] Figure 15 A schematic diagram showing a portion of a channel arranged in a jet device according to some embodiments; and Figure 16A schematic diagram of an exemplary macromonomer is shown according to some embodiments.

[0078] Figures 17A-17C These are bright-field images of hydrogel chambers at various stages of degradation. Figure 17A This is a bright-field image of the hydrogel chamber after incubation for 5 minutes in the presence of a degradation agent. Figure 17B This is a bright-field image of the hydrogel chamber after incubation for 10 minutes in the presence of a degradation agent. Figure 17C This is a bright-field image of the hydrogel chamber after incubation for 20 minutes in the presence of a degradation agent.

[0079] Figure 18 This is a graph showing the degradation time of the hydrogel chamber under different concentrations of degrading agents.

[0080] Figures 19A-19C These are bright-field images of hydrogel chambers photopolymerized using different irradiation times. Figure 19A This is an image of a hydrogel chamber formed by 1 second of irradiation. Figure 19B This is an image of a hydrogel chamber formed using 2-second irradiation. Figure 19C This is an image of a hydrogel chamber formed using 3-second irradiation.

[0081] Figures 20A-20B These are a series of bright-field images of hydrogel chambers that enclose adherent cells. Figure 20A These are a series of images of the hydrogel chambers containing the adherent cells, immediately after the formation of the hydrogel chambers and before the adherent cells adhere to the surface of the jet device. Figure 20B These are a series of images of the hydrogel chambers that enclose adherent cells 72 hours after the formation of the hydrogel chambers.

[0082] Figure 21 This is a schematic diagram showing the photopolymerization and photodegradation of polymer precursors. Detailed Implementation

[0083] Analyzing biological samples at the levels of cellular physical properties, proteomics, transcriptomics, genomics, epigenomics, methylome, secretome, or metabolome allows for the analysis of single cells or cell populations. For example, genetic material from a cell can be analyzed separately or in combination with genetic material from other cells. While analyzing sample components individually can produce higher resolution data with less noise or cross-contamination, this approach can be costly and time-consuming. On the other hand, analyzing a large number of samples or multiple samples can be cost- and time-efficient, but may produce undesirable or less desirable outputs (e.g., due to heterogeneity). For example, specific data may not be traceable to their origin within the sample. Spatial correlations between sample components may also be lost. For instance, two components of a sample may be adjacent to each other in their biologically relevant states, but spatial information between the two components may be lost after aggregating multiple samples or sample components.

[0084] To avoid generating heterogeneous samples, individual components of a sample can be compartmentalized. This allows for the simultaneous or substantially simultaneous processing of individual components of the sample while preserving spatial information and reducing processing steps. Additionally, by preventing cross-contamination and material loss during external sample processing, the resulting data can be of higher quality (e.g., the data can appropriately have a higher signal-to-noise ratio). Some methods preserve spatial information by barcoding individual compartments or groups of compartments within a sample. These methods can combine barcoded components and may not be able to perform multiple determinations of the same component within a compartment or analytical chamber. Some of these methods may require a nucleotide amplification step, which could introduce bias into the generated data.

[0085] To compartmentalize individual components of a biological sample, a polymer matrix (e.g., a hydrogel matrix) can be formed adjacent to or around at least a portion of the individual component within the jet device. The hydrogel matrix can be selectively generated to surround the component after system detection, or it can be generated according to a predefined pattern within the jet device. The hydrogel matrix allows reagents and smaller entities to pass through while maintaining the individual components of the biological sample in place. Because one or more individual components can be positioned within the jet device (e.g., encapsulated), and the positioned components can be exposed to one or more reagents and / or wash solutions during and / or between analyses, multiple assays can be performed within the compartment (e.g., simultaneously, substantially simultaneously, sequentially, etc.).

[0086] This document discloses compositions, systems, and methods for spatially controlled formation of polymer matrices within biological samples. The polymer matrix can be formed by photopolymerization of crosslinkable monomers defined at a location within a jet apparatus. For example, the compositions and systems disclosed herein, by identifying the location of cells and then irradiating a region surrounding the cells to photopolymerize polymer precursors, can be used to form chambers surrounding cells within a jet apparatus, such as in… Figures 1A-1E , Figures 5A-5E , Figures 6A-6F , Figures 8A-8D , Figures 12A-12G , Figures 17A-17C , Figures 19A-19C and Figures 20A-20BThose shown in the diagram. To facilitate this method in various embodiments, the polymer precursor can achieve one or more of the following features. First, the polymer precursor can polymerize rapidly to prevent cell migration from the chamber during polymerization. Second, to facilitate longitudinal cell assays, the polymer precursor can be stable (e.g., maintaining structural fidelity for days, weeks, or months). Third, the polymerized polymer precursor can be biocompatible and, particularly in its pre-polymerized and polymerized forms, exhibit low cytotoxicity. Fourth, the polymer can be stable in the presence of cells; in the presence of an extracellular environment containing oxidizing (e.g., superoxide), reducing (e.g., thiols), and / or extracellular enzymes (e.g., hydrolases, thioredoxins, and / or nucleases); and in the presence of cell-induced mechanical stress. Finally, the polymer can degrade rapidly to facilitate the collection of whole cells, cell secretions, and cell lysates. The polymer precursor of this application fulfills an unmet need by exhibiting one or more combinations of features required for spatially and temporally controlled cell capture, culture, and / or analysis. For various implementations, it has been surprisingly found that multi-arm polymer precursors having crosslinkable functional groups (e.g., acrylates or acrylamides), degradable functional groups (e.g., disulfides or vicinal diols), and oligomeric domains (e.g., polyethylene glycol) on each arm provide one or more features suitable for use with published biological partitioning assays (e.g., photopolymerization chambers to enclose selected cells from biological samples).

[0087] Different measurements can be performed at different locations within the jet apparatus, for example, to test the effects of different treatment conditions. Furthermore, because components are typically not mixed or combined, low component concentrations (e.g., due to dilution) can be prevented. For example, when analyzing genomic material, the amplification step can be avoided due to the preservation of genetic material in each compartment. Interactions between components can also be studied by having two or more components within each compartment. The polymer matrix can be degraded "as needed," enabling controlled localization and release mechanisms. The solution presented herein preserves the spatial information of components and generates data at the cellular, proteomic, transcriptomic, or genomic levels. Because spatial information is preserved, the data can be correlated with phenotypic data (e.g., linked). Moreover, the solution presented herein preserves the spatial information of components and links data at the cellular, proteomic, transcriptomic, or genomic levels (e.g., phenotypic data).

[0088] Although various embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Many variations, modifications, and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various alternatives may be employed with respect to the embodiments of the invention described herein.

[0089] Whenever the term "at least" precedes the first value in a series of two or more values, the term "at least" applies to each value in that series. For example, at least 1, 2, or 3 is equivalent to at least 1, at least 2, or at least 3.

[0090] When the term "less than" precedes the first value in a series of two or more values, the term "less than" applies to each value in that series. For example, less than 3, 2, or 1 is equivalent to less than 3, less than 2, or less than 1.

[0091] As used herein, the terms “connected to,” “connected to,” and “in communication with” generally refer to any form of interaction between two or more entities, including mechanical, electrical, magnetic, electromagnetic, fluid, biological, and thermal interactions. Even if two components are not in direct contact with each other, they may still be connected.

[0092] As used interchangeably herein, the terms “polypeptide” and “peptide” generally refer to a polymer of amino acids, wherein an amino acid is linked to another amino acid via a peptide bond. In some examples, a polypeptide is a protein. Amino acids can be naturally occurring or non-naturally occurring (e.g., amino acid analogs). Polypeptides can be linear or branched. Polypeptides can contain modified amino acids. Polypeptides can be interrupted by non-amino acid groups. Polypeptides can exist as a single chain or associated chains. Polypeptides can contain multiple amino acids. Polypeptides can have secondary and tertiary structures (e.g., a polypeptide can be a protein). In some examples, a polypeptide can contain at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 100, 1,000, 10,000 or more amino acids. Polypeptides can be fragments of larger polymers. In some examples, a polypeptide can be a fragment of a larger polypeptide, such as a fragment of a protein.

[0093] As used herein, the term "amino acid" generally refers to naturally occurring or non-naturally occurring amino acids (e.g., amino acid analogs). Non-naturally occurring amino acids can be engineered or synthetic amino acids.

[0094] As used herein, the term "sample" generally refers to a chemical or biological sample containing biological components. Biological components can include cells, nucleic acids, microbiomes, proteins, cellular combinations, metabolites, combinations thereof, or any other suitable component of a biological sample. For example, a sample can be a biological sample containing one or more cells. As another example, a sample can be a biological sample containing one or more polypeptides. Biological samples can be obtained (e.g., extracted or isolated) from or include blood (e.g., whole blood), plasma, serum, urine, saliva, mucosal excretions, sputum, feces, and tears. Biological samples can be fluid or tissue samples (e.g., skin samples). In some cases, samples can be derived from homogenized tissue samples (e.g., brain homogenate, liver homogenate, or kidney homogenate). In some embodiments, samples can include specific types of cells (e.g., neuronal cells, muscle cells, hepatocytes, or kidney cells). Samples can contain or be obtained from diseased cells or tissues (e.g., tumor cells or necrotic cells). In some embodiments, samples can contain or may be derived from disease-related inclusions (e.g., plaques, biofilms, tumors, or non-cancerous growths). In some embodiments, the sample may include or be available from cell-free bodily fluids, such as whole blood, saliva, or urine. In various embodiments, the sample may include circulating tumor cells. In some cases, the sample may include or be an environmental sample (e.g., soil, waste, or ambient air), an industrial sample (e.g., a sample from any industrial process), or a food sample (e.g., dairy products, vegetable products, or meat products). The sample may be treated before being loaded into the microfluidic device. For example, the sample may be treated to purify a cell type or peptide and / or contain reagents.

[0095] As used herein, the term "polymer matrix" generally refers to a phase material (e.g., a continuous phase material) comprising at least one polymer. In some embodiments, a polymer matrix refers to at least one polymer and interstitial spaces not occupied by the polymer. A polymer matrix may be composed of one or more types of polymers. A polymer matrix may comprise linear, branched, and cross-linked polymer units. A polymer matrix may also comprise non-polymer species inserted into its interstitial spaces not occupied by polymer chains. The inserted species may be solid, liquid, or gaseous species. For example, the term "polymer matrix" may encompass dry hydrogels, hydrated hydrogels, and hydrogels containing glass fibers. A polymer matrix may comprise polymer precursors, which generally refer to one or more molecules that can trigger or initiate a polymerization reaction upon activation. Polymer precursors may be activated by electrochemical energy, photochemical energy, photons, magnetic energy, or any other suitable energy.

[0096] As commonly used herein, the terms "viability" or "cell viability" or "viability percentage" refer to the percentage of live, healthy cells within a population, typically expressed as a ratio or percentage. In some embodiments, viability is a measure of the ability of cells to survive and function as a function of the total number of cells in a given sample. In some embodiments, evaluating cell viability provides insight into the health, viability, and overall function of a cell population under specific conditions or treatments. In some embodiments, techniques such as staining with specific dyes or using assays are employed to assess cell viability in a laboratory setting.

[0097] The diverse range of photosynthetic and biodegradable gels discussed below can be used to implement the systems and methods described herein. Guidance on selecting such gels for desired properties (including but not limited to biocompatibility, gelation rate, degradation rate, etc.) is provided in the following references, which are incorporated herein by reference: Kharkar et al., Chem. Soc. Rev., 42: 7335-7372 (2013); Kharkar et al., Polymer Chem., 6(31): 5565-5574 (2015); Neumann et al., Acta Biomater., 39: 1-11 (2016); DeForest et al., Nature Chemistry, 3(12): 925-931 (2012); Bowman et al., US Patent 9631092; LeValley et al., ACS Appl. Bio. Mater., 3(10): 6944-6958 (2020); Kabb et al., ACS Appl. Mater. Interfaces, 10: 16793-16801 (2018); Fairbanks et al., Macromolecules, 44: 2444-2450 (2011); Fairbanks et al., Adv. Mater., 21(48): 5005-5010 (2009); Sugiura et al., US Patent Publication US2016 / 0177030; Shih et al., Biomacromolecules, 13(7): 2003-2012 (2012); etc. In some embodiments, the photosynthetic gel is formed using a photoinitiator for free radical polymerization. In some embodiments, the photoinitiator includes Irgacure 2959, lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), or eosin Y (see, for example, Choi et al., Biotechniques, 66(1): 40-53 (2019)). In some embodiments, the hydrogel precursor includes hyaluronic acid, chitosan, heparin, alginate, polyethylene glycol (PEG), multi-arm PEG, poly(ethylene glycol)-b-poly(propylene oxide)-b-poly(ethylene glycol) (PEG-PPO-PEG), poly(lactic acid-co-glycolic acid)-b-poly(ethylene glycol)-b-poly(lactic acid-co-glycolic acid) (PLGA-PEG-PLGA), and poly(vinyl alcohol). In some embodiments, the polymer precursor includes PEG or multi-arm PEG. In some embodiments, the polymer precursor includes an enzymatically degradable crosslinking agent. In some embodiments, such an enzymatically degradable crosslinking agent is degraded by esterases or peptidases.In some embodiments, the polymer precursor comprises a photodegradable crosslinking agent. In some embodiments, such a photodegradable crosslinking agent comprises a nitrobenzyl group. In some embodiments, such a photodegradable crosslinking agent comprises a coumarin moiety. In some embodiments, the photodegradable hydrogel is used in conjunction with the methods described herein, for example, because the photodegradation of the hydrogel chambers is selective and on-demand, allowing specific hydrogel chambers to degrade without affecting the unselected hydrogel chambers being unaffected. In some embodiments, the hydrogel chambers undergo non-selective degradation, such that all hydrogel chambers in a given channel (or other container) degrade simultaneously. In some embodiments, such non-selective degradation is performed using a cleaving agent that specifically cleaves unstable bonds in the hydrogel. For example, such cleaving agents include reducing agents. In some embodiments, such non-specific degradation is performed using an enzyme that cleaves bonds or chemical elements in the hydrogel. For example, chemical elements include, but are not limited to, peptides, polysaccharides, and oligonucleotides.

[0098] Figure 14A A portion of the system as presented herein is shown, including the energy source. Figure 14B The polymer matrix formed around the biological components in a part of the system provided herein is shown. Figure 14C A method for forming a polymer matrix around a biological component in a system as provided herein is shown. Figure 15 A schematic diagram showing a portion of the channels arranged in the jet device is shown.

[0099] The channels or chambers of the jet device can receive or be configured to receive biological samples. Figure 15 A schematic diagram showing a portion of channel 100, which may be disposed in at least a portion of a jetting apparatus as provided herein. The jetting apparatus may include channel 100. Channel 100 may include a first surface 101. Furthermore, channel 100 may include a second surface 102. In some embodiments, the first surface 101 and the second surface 102 are disposed, placed, or positioned opposite each other (e.g., as shown in the diagram). Figure 15 (As depicted in the figures). In some embodiments, the first surface 101 may be the lower surface. In some embodiments, the second surface 102 may be the upper surface. The terms "lower" and "upper" are not intended to be limiting and are used herein for convenience when referring to the accompanying drawings. Channel 100 may receive a biological sample containing one or more biological components 50, 51. Channel 100 may receive one or more polymer precursors. Figure 15As shown, biological components 50 and 51 may include cells. However, as discussed herein, biological components may include tissues, proteins, nucleic acids, etc. In some embodiments, the first surface 101, the second surface 102, or both surfaces may be coupled to or receive, or configured to coupled to or receive at least one of one or more biological components 50 and 51. In some cases, the first surface 101 may be coupled to or receive, or configured to coupled to or receive biological components (e.g., biological components 50 and 51). In some cases, the second surface 102 may be coupled to or receive, or configured to coupled to or receive biological components (e.g., biological components 50 and 51).

[0100] In some cases, channels may have rectangular, circular, semi-circular, oval, or other suitable cross-sections. Therefore, a channel may have a single inner surface. In other cases, channels may have triangular, square, rectangular, polygonal, or other cross-sections. Therefore, a channel may have three or more inner surfaces. One or more inner surfaces may be connected to or receive, or configured to connect to or receive, one or more biological components.

[0101] In some cases, the first surface 101, the second surface 102, or both surfaces 101, 102 may be functionalized, for example, with a coating (e.g., a surface coating). In some embodiments, the surface coating may be a surface polymer. Some non-limiting examples of the surface coating may include a trapping agent (e.g., pyridine formaldehyde (PCA)), a functional group that traps one or more portions (e.g., chemical portions), acrylamide, agarose, biotin, streptavidin, strep-tag II, a linker, an aldehyde-containing functional group, phosphate ester, silicate ester, ester, acid, amide, alkyne, azide, aldehyde dithiopentane, or combinations thereof. In various embodiments, the surface coating may contain a functional group that traps one or more portions. For example, acrylamide, agarose, etc., may contain such a functional group. In some embodiments, the surface polymer may contain polyethylene glycol (PEG), thiols, olefins, alkynes, azides, or combinations thereof. In various embodiments, the surface polymer may contain a silane polymer. In some implementations, the surface polymer may be functionalized with at least one of oligonucleotides, antibodies, cytokines, chemokines, proteins, antibody derivatives, antibody fragments, carbohydrates, toxins, or aptamers.

[0102] In some cases, the first surface 101, the second surface 102, or both surfaces 101, 102 may contain one or more barcodes (e.g., nucleic acid barcodes). In some embodiments, the first surface 101, the second surface 102, or both surfaces 101, 102 may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 100, 200, 300, 400, 500, 600, 700, 1,000, 10,000, 50,000, 100,000, 250,000, 500,000, 1,000,000, 2,000,000, 5,000,000, 10,000,000, 15,000,000 barcodes or any number of barcodes between any two numbers mentioned herein. Barcodes can cover approximately 500 nm. 2 to approximately 500 nm 2 The area. In some embodiments, the first surface 101, the second surface 102, or both surfaces 101, 102 may include a total of up to about 10,000,000 barcodes. The barcodes may be different from each other (e.g., each barcode may be unique). In some embodiments, a first portion or subset of the barcodes may be different from a second portion or subset of the barcodes. There may be 2, 3, 4, 5, 10, 15, 20, 25, 50, 75, 100, 1,000, 10,000 barcode portions or subsets, or any number of barcode portions or subsets between any two numbers mentioned herein. In some cases, the barcode (or a portion / subset of the barcode) may be associated with the barcode's location on the surface (location coordinates on the channel surface (e.g., x-coordinate, y-coordinate)). The barcode may be attached or coupled to the captured biological component. In some embodiments, the barcode may be a unique identifier that distinguishes a biological component from other biological components (e.g., identifying a first biological component relative to a second biological component). In some embodiments, the barcode may contain the captured biological component or a nucleic acid sequence (e.g., a common sequence) used in amplification. In some embodiments, the barcode may contain a unique identifier, which may include a unique nucleic acid sequence (e.g., a DNA sequence, RNA sequence, etc.), a protein tag, an antibody, or an aptamer. In some embodiments, the barcode may contain a fluorescent molecule. In some embodiments, the location of the captured biological component may be associated with the unique identifier to, for example, preserve spatial information about the biological component.

[0103] In some embodiments, the jet device may be a flow cell. For example, the jet device may be used for sequencing (e.g., DNA or RNA sequencing). In some embodiments, the jet device may be a microjet device. In some embodiments, the jet device may be a nanojet device.

[0104] The systems disclosed herein may include one or more energy sources. The energy source may be in communication with a jetting device. In some cases, the energy source may be used to form one or more polymer matrices within the jetting device (e.g., on or adjacent to a channel or chamber of the jetting device). In some embodiments, the energy source may include a photogenerating device, a thermal generating device, an electrochemical reaction generating device, an electrode, or a microwave device. The polymer matrix may be formed within a channel of the jetting device. The energy source may direct or transfer energy to a predetermined location within the jetting device. The energy may induce or activate one or more polymer precursors to form a polymer matrix (e.g., polymerization) at the predetermined location.

[0105] In some embodiments, the polymer matrix may comprise a hydrogel. In some embodiments, the hydrogel may be sufficiently porous, or have pores of suitable size, to allow reagents (e.g., enzymes, compounds, small molecules, antibodies, etc.) to move or transfer through the polymer matrix, while the hydrogel may not allow biological components (e.g., DNA, RNA, proteins, cells, etc.) to move or transfer through the polymer matrix. In some embodiments, the pore diameter may be from 5 nm to 100 nm. In some embodiments, the pore diameter may be from 5 nm to 10 nm, 10 nm to 20 nm, 20 nm to 30 nm, 30 nm to 40 nm, 50 nm to 60 nm, 60 nm to 70 nm, 70 nm to 80 nm, 80 nm to 90 nm, or 90 nm to 100 nm. In some embodiments, the pore diameter may be greater than 100 nm. In some embodiments, the pore diameter may be less than 5 nm. Reagents may include enzymes or primers less than 50 base pairs (bp) in size. Primers may comprise single-stranded DNA (ssDNA). In some embodiments, the primer size can be from 5 bp to 50 bp. In some embodiments, the primer size can be from 5 bp to 10 bp, 10 bp to 20 bp, 20 bp to 30 bp, 30 bp to 40 bp, or 40 bp to 50 bp. In some embodiments, the primer size can exceed 50 bp. In some cases, the primer size can be less than 5 bp. Reagents may include lysozyme, proteinase K, hexamer (e.g., random hexamer), polymerase, transposase, ligase, catalytic enzyme, deoxyribonuclease, deoxyribonuclease inhibitor, ribonuclease, ribonuclease inhibitor, DNA oligomer, deoxyribonucleotide triphosphate, buffer, detergent, salt, divalent cation, or any other suitable reagent.

[0106] In some embodiments, the polymer matrix may comprise a hydrogel. In some embodiments, the hydrogel may be sufficiently porous, or have pores of suitable size, to allow reagents (e.g., enzymes, compounds, small molecules, antibodies, etc.) to move or transfer through the polymer matrix, while the hydrogel may not allow biological components (e.g., DNA, RNA, proteins, cells, etc.) to move or transfer through the polymer matrix. In some embodiments, the pore diameter may be from 5 nm to 100 nm. In some embodiments, the pore diameter may be from 5 nm to 10 nm, 10 nm to 20 nm, 20 nm to 30 nm, 30 nm to 40 nm, 50 nm to 60 nm, 60 nm to 70 nm, 70 nm to 80 nm, 80 nm to 90 nm, or 90 nm to 100 nm. In some embodiments, the pore diameter may be greater than 100 nm. In some embodiments, the pore diameter may be less than 5 nm. Reagents may include enzymes or primers less than 50 base pairs (bp) in size. Primers may comprise single-stranded DNA (ssDNA). In some embodiments, the primer size can be from 5 bp to 50 bp. In some embodiments, the primer size can be from 5 bp to 10 bp, 10 bp to 20 bp, 20 bp to 30 bp, 30 bp to 40 bp, or 40 bp to 50 bp. In some embodiments, the primer size can exceed 50 bp. In some cases, the primer size can be less than 5 bp. Reagents may include lysozyme, proteinase K, hexamer (e.g., random hexamer), polymerase, transposase, ligase, catalytic enzyme, deoxyribonuclease, deoxyribonuclease inhibitor, ribonuclease, ribonuclease inhibitor, DNA oligomer, deoxyribonucleotide triphosphate, buffer, detergent, salt, divalent cation, or any other suitable reagent.

[0107] Figure 14A As shown in this document, this is part of a system including energy source 203. Figure 14A The implementation scheme may include aspects similar to Figure 15 Components of a component. For example. Figure 14A The implementation scheme includes those that can be similar to Figure 15 Channel 100 and Channel 200. It should be understood that the illustrated embodiments may have similar characteristics. Therefore, similar characteristics are indicated by similar reference numerals, where the leading digits increment to "2". Therefore, the relevant disclosures regarding the characteristics of similarity identification described above will not be repeated below. Furthermore, the specific characteristics of the system provided herein and... Figure 14AThe relevant components shown may not be indicated or labeled by reference numerals in the accompanying drawings or specifically discussed in the following written description. However, such features may be explicitly identical or substantially identical to features depicted and / or described with respect to other embodiments. Therefore, the relevant descriptions of such features are equally applicable. Figure 14A The characteristics of the system and related components. Regarding... Figure 15 Any suitable combination and variation of the characteristics described in the system and component diagrams can be used with Figure 14A The systems and components are used together, and vice versa. This disclosure also applies to other embodiments depicted in the following figures and described below.

[0108] Continue to refer to Figure 14A The system channel 200 may include a first surface 201 and a second surface 202. In some embodiments, the energy source 203 may include one or more energy emitting portions (e.g., energy emitting portion 205). In some embodiments, the energy source 203 may include one or more non-emitting portions (e.g., non-emitting portion 204). The non-emitting portion 204 may not emit or may be configured not to emit energy. In some embodiments, the emitting portion 205 may emit energy to at least a portion of the jet device in the form of electromagnetic waves (e.g., microwaves, light, heat, etc.). In some embodiments, the emitting portion 205 may emit energy to the jet device. In some embodiments, the jet channel may be coupled to or arranged on a movable stage. In other embodiments, light may be projected onto at least a portion of or onto the jet channel to generate one or more polymer matrices. Light may be directed to various components of the jet channel. In some embodiments, the emitting portion 205 may be coupled to an objective lens (e.g., a microscope objective lens), wherein the objective lens may be movable to different portions of the jet device. Objective lenses can provide shapes (e.g., virtual or physical masks) that allow light to pattern on the jetting device to form a polymer matrix similar to or complementary to the pattern. In various embodiments, one or more polymer precursors in or mixed with the biological sample in the jetting device can absorb the emitted energy 206. In some embodiments, the emitted energy 206 can be, or is sufficient to, form a polymer matrix from one or more polymer precursors. For example, a portion of one or more polymer precursors within the channel 200 of the jetting device can be activated by the emitted energy and can initiate a polymerization reaction to form a polymer matrix.

[0109] In some embodiments, an energy source can emit energy to a large portion or almost the entire surface of the jet channel. A physical mask can be used to block energy emitted to one or more portions of the jet channel. The energy source (e.g., a light source) can be coupled to the jet device via an objective lens (e.g., a microscope objective or lens). The energy source can be directed to a portion of the jet channel (e.g., via a movable objective lens). In some cases, the light source, objective lens, and / or jet channel are movable to allow energy to be emitted into the jet channel to generate a pattern on at least a portion of the surface of the jet device. The polymer matrix can be formed similarly to or complementary to the pattern emitted by the energy.

[0110] The polymer precursor may contain an activating molecule that can absorb emitted energy 206 to initiate polymerization of one or more polymer precursors in a jet apparatus. Non-limiting examples of the activating molecule may include photocatalysts, photoactivators, photoacid generators, or photobase generators. In some embodiments, a first polymer matrix 208 and / or a second polymer matrix 209 may be formed on or adjacent to the biological component 50. In some embodiments, the first polymer matrix 208 and the second polymer matrix 209 may form an analytical chamber or compartment 220 that separates (e.g., physically separates) the biological component 50 from other biological components (e.g., biological components 51, 52, or 53) in the jet apparatus. In other words, the polymer matrix may compartmentalize channels (e.g., channel 200). In various embodiments, the polymer matrix may partially surround the biological component. For example, the polymer structure surrounding the biological component may form a closed structure (e.g., a hollow cylindrical polymer structure) or a partially open structure (e.g., a crescent-shaped polymer structure). In some embodiments, two or more polymer matrices may be formed adjacent to the biological component to form compartments that separate the biological component from other biological components. In some embodiments, the polymer matrix may include or form walls (e.g., polymer matrix walls).

[0111] In various embodiments, the polymer matrix comprises a hydrogel. In some embodiments, the polymer matrix wall may be a hydrogel wall. In some embodiments, the hydrogel or hydrogel wall may comprise polyethylene glycol (PEG)-thiol, PEG-acrylate, acrylamide, N,N'-bis(acryloyl)cysteine ​​(BAC), PEG, polypropylene oxide (PPO), polyacrylic acid, poly(hydroxyethyl methacrylate) (PHEMA), poly(methyl methacrylate) (PMMA), poly(N-isopropylacrylamide) (PNIPAAm), poly(lactic acid) (PLA), polylactic acid-glycolic acid copolymer (PLGA), polycaprolactone (PCL), poly(vinyl sulfonic acid) (PVSA), etc. (L-Aspartic acid), poly(L-glutamic acid), polylysine, agar, agarose, alginate, heparin, alginate sulfate, dextran sulfate, dextran-acrylamide, hyaluronic acid, pectin, carrageenan, gelatin, chitosan, cellulose, collagen, bisacrylamide, diacrylate, diallylamine, triallylamine, divinyl sulfone, diethylene glycol diallyl ether, ethylene glycol diacrylate, polymethylene diacrylate, polyethylene glycol diacrylate, trimethylolpropane trimethacrylate, ethoxylated trimethylolpropane triacrylate, ethoxylated pentaerythritol tetraacrylate, or combinations or mixtures thereof. The hydrogel or hydrogel wall may contain a biodegradable crosslinking agent (e.g., N,N'-bis(acryloyl)cysteine, chitosan, poly(ε-caprolactone) diacrylate, polylactide diacrylate, polylactide dimethacrylate, poly(lactide-glycolic acid) copolymer, polycaprolactone molecules or other suitable biodegradable crosslinking agents).

[0112] In some embodiments, the surface of the polymer matrix or hydrogel can be functionalized by coupling functional groups to the polymer matrix or hydrogel. Some non-limiting examples of functional groups may include trapping reagents (e.g., pyridine carboxaldehyde (PCA)), acrylamide, agarose, biotin, streptavidin, strep-tag II, linkers, functional groups comprising aldehydes, phosphate esters, silicates, esters, acids, amides, aldehyde dithiopentane, PEG, thiols, alkenes, alkynes, azides, or combinations thereof. In some cases, the functionalized polymer matrix can be used to trap biomolecules inside a polymer matrix compartment formed adjacent to (e.g., around or on) a biological component. Biomolecules can be produced by the biological component (e.g., from the secretome of cells). The functionalized surface of the polymer matrix inside the compartment can be used to trap reagents or molecules from outside the compartment. The functionalized surface can increase the surface area covered by reagents, molecular sensors, or any molecules of interest (e.g., antibodies).

[0113] In some embodiments, the compartments surrounding the biological component may include a polygonal base. In various embodiments, the compartments surrounding the biological component may include a circular or oval base (see, for example...). Figure 14C (Cell 220 or cell 222 in the cell). In some embodiments, the thickness (e.g., width) of the polymer matrix wall of the cell can be from 1 μm to 250 μm. The thickness of the polymer matrix wall or compartment can be 1 μm to 5 μm, 1 μm to 10 μm, 1 μm to 20 μm, 1 μm to 30 μm, 1 μm to 40 μm, 1 μm to 50 μm, 1 μm to 100 μm, 1 μm to 150 μm, 1 μm to 250 μm, 5 μm to 10 μm, 5 μm to 20 μm, 5 μm to 30 μm, 5 μm to 40 μm, 5 μm to 50 μm, 5 μm to 100 μm, 5 μm to 150 μm, 5 μm to 250 μm, 10 μm to 20 μm, 10 μm to 30 μm, 10 μm to 40 μm, 10 μm to 50 μm, 10 μm to 100 μm, 10 μm to 150 μm, 10 μm to 250 μm, 20 μm to 30 μm, 20 μm to 40 μm, 20 μm to 50 μm, 20 μm to 100 μm, 20 μm to 150 μm, 20 μm to 250 μm, 30 μm to 40 μm, 30 μm to 50 μm, 30 μm to 100 μm, 30 μm to 150 μm, 30 μm to 250 μm, 40 μm to 50 μm, 40 μm to 100 μm, 40 μm to 150 μm, 40 μm to 250 μm, 50 μm to 100 μm, 50 μm to 150 μm, 50 μm to 250 μm, 100 μm to 150 μm, 100 μm to 250 μm or 150 μm to 250 μm. The thickness of the polymer matrix walls or compartments can be approximately 1 μm, approximately 5 μm, approximately 10 μm, approximately 20 μm, approximately 30 μm, approximately 40 μm, approximately 50 μm, approximately 100 μm, approximately 150 μm, or approximately 250 μm. The thickness of the polymer matrix walls or compartments can be at least 1 μm, 5 μm, 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 100 μm, 150 μm, 250 μm, or greater. The thickness of the polymer matrix walls or compartments can be at most 5 μm, 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 100 μm, 150 μm, or 250 μm. The thickness of the polymer matrix walls or compartments can be less than 1 μm.

[0114] Continue to refer to Figure 14APolymer matrices 208, 209, or at least a portion thereof, may be attached to the first surface 201, the second surface 202, or both surfaces 201, 202. In some embodiments, the polymer matrix or at least a portion thereof may be suitably attached to a third surface, a fourth surface, a fifth surface, etc. In various embodiments, polymer matrices 208, 209 may extend from the first surface 201 to the second surface 202 (e.g., through at least a portion of the cavity of the channel 200 or chamber), such that the polymer matrix surrounds or substantially surrounds the biological component 50. In some embodiments, two or more biological components that are physically very close together (e.g., Figure 14C The biological components 50, 51 can be separated (e.g., by stirring or agitating the jet device). The jet device can be stirred or agitated by physical movement, the use of acoustic pulses, alteration of flow in the channel, or any other suitable stirring method. A polymer matrix can then be formed around (or partially around) the separated biological components. Figure 14B The polymer matrix 208, 209 formed around the biological component 50 after separation from the biological component 51 is shown. Figure 14C This illustrates the process of separating two very close biological components 50 and 51 according to various embodiments. That is, biological components 50 and 51 can be separated by stirring or agitating the jet device. In some embodiments, the separation of biological components is achieved by fluid pressure, flow pulsation, dielectrophoresis, photothermal flow, or some combination thereof. In some cases, the separation of biological components is achieved by acoustic vibration. Figure 14C It also shows that after separating the biological components 50 and 51, a polymer matrix is ​​formed around the biological component 50 to generate compartment 222.

[0115] Continue to refer to Figure 14AIn some cases, energy source 203 may be configured to form or generate one or more emitting portions 205 and one or more non-emitting portions 204. The systems disclosed herein may also include spatial energy modulation elements to direct energy from the energy source to one or more target portions of the jet apparatus. For example, the spatial energy modulation element may be configured to selectively direct energy from the energy source to form a polymer matrix on or adjacent to at least a portion of the biological component. The spatial energy modulation element may be configured to selectively direct energy by, for example, inhibiting or preventing energy from being directed to one or more portions other than one or more target portions of the jet apparatus. In some embodiments, the spatial energy modulation element may include a physical mask. In some cases, the spatial energy modulation element may include a virtual mask. In some cases, the spatial energy modulation element may be configured to control one or more electrodes that can selectively provide energy to one or more target portions of the jet apparatus. The electrode concept may also be used to provide spatially modulated energy to form a hydrogel structure. In some specific embodiments, one or more electrodes may be arranged at predetermined locations in the jet channel, thus allowing hydrogel formation in those locations. In alternative embodiments, the electrodes may be in the form of an array. The elements of the array can be turned on or off as needed to generate the desired energy spatial pattern and form the desired hydrogel shape. For example, one or more electrodes (e.g., an electrode array) can be disposed within one or more portions of the jet device. As another example, one or more electrodes (e.g., an electrode array) can be in communication (e.g., electrical communication) with one or more portions of the jet device.

[0116] In some embodiments, the mask may prevent or be configured to prevent one or more portions of the energy emitting surface 210 of the energy source 203 (e.g., non-emitting portion 204) from emitting energy. In some embodiments, the mask may be a virtual mask (e.g., computer code or a digital system). In some embodiments, the mask may prevent energy emission to a location where the biological component is present. This may allow or permit the formation of a polymer matrix adjacent to, on, or encapsulating the biological component (e.g., to retain cells, proteins, DNA molecules, RNA molecules, or other target molecules at a location on the jet channel). In other embodiments, the mask may facilitate polymerization such that the polymer matrix is ​​on the biological component. In various embodiments, the mask may be a physical mask (e.g., an opaque material, thermal shield, or electromagnetic shield). In some embodiments, a detector or combination thereof for detecting or identifying the location of the biological component may be used to generate the mask (e.g., a virtual mask or a physical mask). In some embodiments, the detector includes a camera. In some embodiments, the detector includes a photodetector, a conductivity detector, an ultrasonic detector, an ultrasonic sensor, a piezoelectric sensor, a combination thereof, or another suitable detection device.

[0117] In some embodiments, the first surface 201 or the second surface 202 may include a detector that detects or is configured to detect one or more locations of one or more biological components in the jet apparatus (e.g., in channel 200). In some embodiments, the energy source 203 may include, be coupled to, or communicate with a detector that detects or is configured to detect the location of biological components in the jet apparatus. In various embodiments, an image obtained from at least a portion of the jet apparatus may be used to generate a mask. The mask may allow or permit the energy source 203 to emit energy at or to one or more locations of one or more biological components on or adjacent to the first surface 201. The mask may inhibit or prevent the energy source 203 from emitting energy at or to one or more locations of one or more biological components on or adjacent to the first surface 201. In some embodiments, the image may be obtained from a camera (e.g., a digital camera, a fluorescence imaging camera, etc.). In some embodiments, the camera may be coupled to, connected to, or communicated with the energy source 203. For example, a camera (not shown) may be electrically connected to the energy source 203. In some embodiments, the energy source 203 may include a camera. In various embodiments, the energy source 203 may include a microscope (e.g., a fluorescence microscope, a confocal microscope, a lensless imaging system, a transmission electron microscope (TEM), a scanning electron microscope (SEM), etc.). The microscope may be used to detect one or more locations of one or more biological components (e.g., in combination with a detector).

[0118] This disclosure provides polymer precursors. In some embodiments, the polymer precursor comprises an oligomeric domain. In some embodiments, the oligomeric domain comprises three or more arms. In some embodiments, each arm of the oligomeric domain comprises a degradable functional group and a crosslinkable functional group. In some embodiments, the crosslinkable functional group of one of the three or more arms is configured to crosslink with another crosslinkable functional group of another polymer precursor in response to a first stimulus, thereby obtaining a polymeric form of the polymer precursor. In some embodiments, the degradable functional group is configured to dissolve the polymeric form of the polymer precursor in response to a second stimulus being cleaved. In some cases, the degradable functional group is configured to dissolve the polymeric form of the monomer in response to a second stimulus being cleaved. In some cases, the oligomeric domain comprises four or more arms. In some cases, the polymer precursor comprises a monomer selected from structures (I), (II), (IV)-(VI), (XII), (XIII), (XIV)-(XVIII), (XX), and combinations thereof.

[0119] Surprisingly, it has been found that including degradable and crosslinkable functional groups on each arm of the polymer precursor can improve its suitability for cell encapsulation methods. Such methods impose numerous requirements, including rapid spatially and temporally controlled polymerization; rapid spatially and temporally controlled degradation; controllable porosity; biocompatibility; and sufficient rigidity to withstand the mechanical stresses of cell adhesion. In some cases, each arm contains examples of degradable and crosslinkable functional groups. In some cases, each arm contains examples of oligomeric domains. In some cases, each arm contains polyethylene glycol units. In some cases, each polyethylene glycol unit contains between about 1 and 500, about 1 and 200, about 1 and 100, about 1 and 50, or about 20 and 50 polyethylene glycol units. Therefore, each arm can include approximately 250 to 500 Daltons, approximately 250 to 1000 Daltons, approximately 250 to 2500 Daltons, approximately 250 to 5000 Daltons, approximately 250 to 10000 Daltons, approximately 250 to 15000 Daltons, approximately 250 to 25000 Daltons, approximately 500 to 1000 Daltons, approximately 500 to 25000 Daltons, approximately 500 to 5000 Daltons, approximately 500 to 10000 Daltons, approximately 500 to 15000 Daltons, approximately 500 to 25000 Daltons, approximately 1000 to 2500 Daltons, approximately 1000 to 5000 Daltons. Molecules ranging from approximately 1,000 to 10,000 Daltons, approximately 1,000 to 15,000 Daltons, approximately 1,000 to 25,000 Daltons, approximately 2,500 to 5,000 Daltons, approximately 2,500 to 10,000 Daltons, approximately 2,500 to 15,000 Daltons, approximately 2,500 to 25,000 Daltons, approximately 5,000 to 10,000 Daltons, approximately 5,000 to 15,000 Daltons, approximately 5,000 to 25,000 Daltons, or approximately 15,000 to 25,000 Daltons. In certain cases, each arm may contain a molecular weight of approximately 500 to 2,500 or 500 to 5,000 Daltons.Similarly, the polymer precursor may include approximately 1,000 to 2,000 Daltons, approximately 1,000 to 10,000 Daltons, approximately 1,000 to 20,000 Daltons, approximately 1,000 to 40,000 Daltons, approximately 1,000 to 60,000 Daltons, approximately 1,000 to 100,000 Daltons, approximately 2,000 to 40,000 Daltons, approximately 2,000 to 6,000 Daltons, approximately 2,000 to 10,000 Daltons, approximately 2,000 to 20,000 Daltons, approximately 2,000 to 40,000 Daltons, approximately 2,000 to 6,000 Daltons. Molecules ranging from 0 Daltons to approximately 2,000 to 100,000 Daltons, approximately 10,000 to 20,000 Daltons, approximately 10,000 to 40,000 Daltons, approximately 10,000 to 60,000 Daltons, approximately 10,000 to 100,000 Daltons, approximately 20,000 to 40,000 Daltons, approximately 20,000 to 60,000 Daltons, approximately 20,000 to 100,000 Daltons, approximately 40,000 to 10,000 Daltons, or approximately 60,000 to 100,000 Daltons. In specific cases, polymer precursors may have molecular weights ranging from approximately 2,000 to 10,000 or approximately 2,000 to 20,000 Daltons. In specific cases, each arm may be equivalent. Examples of such structures include structures (I), (II), (IV)-(VI), (XII), (XIII), (XIV)-(XVIII), (XX), and in. Figure 16 The broad chemical structure presented in it.

[0120] In many cases, the polymer precursor is the only polymerizable species in the hydrogel formulation. More broadly, in some cases, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 98%, and at least about 99% of the polymerizable groups (e.g., crosslinkable functional groups, such as acrylamide, acrylate, etc.) in the formulation are composed of one type of polymer precursor structure (e.g., any of structures (I)-(XX)) relative to all polymerizable groups from one or more types of polymer precursor structures present in the formulation.

[0121] In some cases, degradable functional groups include disulfides, β-thioether esters, amide hydroxymethyl and vicinal diols, vicinal diols, alginate backbones, dextran backbones, chitosan backbones, hyaluronic acid backbones, chondroitin sulfate backbones, or carboxymethyl cellulose backbones, or combinations thereof.

[0122] In some cases, the degradable functional groups include disulfides. After polymer matrix synthesis, the polymer matrix can be cleaved using thiols (such as glutathione or β-mercaptoethanol), disulfides (such as glutathione disulfide), or reducing agents (such as TCEP or dithiothreitol). Such systems offer the advantage of cleavage under mild, biologically compatible conditions, while simultaneously maintaining stability in the absence of disulfide degrading agents.

[0123] In some cases, degradable functional groups include vicinal diols. Compared to hydrolyzable and reductively cleavable functional groups, vicinal diols offer the advantage of enhanced stability, allowing their use under harsh conditions and with cells activated in polymer cleavage. Many cells secrete reactive species capable of degrading reactive functional groups. For example, some neutrophils and macrophages generate the oxidant superoxide extracellularly, while many adherent cells secrete free thiols and regulate extracellular thiol and disulfide concentrations to mediate intercellular signaling. Reactive degradable functional groups such as disulfides and sugars can therefore be unstable in the presence of certain cell types and can exhibit non-negligible background degradation rates in the presence of these cells. The degradable functional groups of vicinal diols often address this issue by being non-reactive to enzymes and molecules secreted by cells. At the same time, vicinal diols are readily cleavable with mild, biocompatible oxidants such as periodate, permanganate, and peroxides in the presence of manganese catalysts. Polymer matrices formed from monomers containing vicinal diols can therefore be formed and degraded rapidly, as disclosed, for example, in Examples 9 and 10.

[0124] Examples of porogens consistent with this disclosure include particles (e.g., polymers, ceramics, metals, metal oxides, or hydrogel particles), polymers (such as polyethylene glycol and alginate), and vesicles (such as liposomes or micelles). In some embodiments, the porogen includes poly(ethylene glycol). The size, concentration, and type of porogen included in the polymer precursor can be selected based on the desired polymer matrix porosity.

[0125] Figure 16 A schematic diagram of an exemplary macromonomer is shown. In some embodiments, the macromonomer comprises a hydrophilic water group 1601, a crosslinkable functional group 1602, a degradable functional group 1603, or a combination thereof. In some cases, the hydrophilic water group comprises an oligomer, such as polyethylene glycol.

[0126] In some embodiments, the oligomer domain comprises four or more arms. In some embodiments, the oligomer domain is hydrophilic. In some embodiments, the oligomer domain includes poly(ethylene glycol), poly(N-vinylpyrrolidone), poly(acrylic acid), poly(methacrylic acid), poly(vinyl alcohol), poly(L-lysine), poly(2-ethyl-2-oxazoline), poly(maleic acid), poly(vinyl phosphate), poly(acrylamide), poly(vinylamine), poly(ethylene oxide-co-propylene oxide), poly(N-isopropylacrylamide), poly(vinylphosphonic acid), poly(2-vinyl-1-methylpyridinium bromide), poly(N,N-diethylacrylamide), poly(N,N-dimethylacrylamide), poly(2-hydroxyethyl acrylate), poly(2-hydroxy methacrylate-co-methacrylic acid), poly(propylene oxide), poly(2-(dimethylamino)ethyl methacrylate), or poly(N-(2-hydroxypropyl)methacrylamide) or combinations thereof. In some embodiments, the oligomer domain includes poly(ethylene glycol). In some embodiments, the oligomer domain includes poly(N-vinylpyrrolidone). In some embodiments, the oligomer domain includes poly(acrylic acid). In some embodiments, the oligomer domain includes poly(methacrylic acid). In some embodiments, the oligomer domain includes poly(vinyl alcohol). In some embodiments, the oligomer domain includes poly(L-lysine). In some embodiments, the oligomer domain includes poly(2-ethyl-2-oxazoline). In some embodiments, the oligomer domain includes poly(maleic acid). In some embodiments, the oligomer domain includes poly(vinyl phosphate). In some embodiments, the oligomer domain includes poly(acrylamide). In some embodiments, the oligomer domain includes poly(vinylamine). In some embodiments, the oligomer domain includes poly(ethylene oxide-co-propylene oxide). In some embodiments, the oligomer domain includes 1 to 500 monomer units (e.g., PEG). 500 ).

[0127] In some embodiments, the oligomer domain includes poly(N-isopropylacrylamide). In some embodiments, the oligomer domain includes poly(vinylphosphonic acid). In some embodiments, the oligomer domain includes poly(2-vinyl-1-methylpyridinium bromide). In some embodiments, the oligomer domain includes poly(N,N-diethylacrylamide). In some embodiments, the oligomer domain includes poly(N,N-dimethylacrylamide). In some embodiments, the oligomer domain includes poly(2-hydroxyethyl acrylate). In some embodiments, the oligomer domain includes poly(2-hydroxy methacrylate-co-methacrylic acid). In some embodiments, the oligomer domain includes poly(propylene oxide). In some embodiments, the oligomer domain includes poly(2-(dimethylamino)ethyl methacrylate). In some embodiments, the oligomer domain includes poly(N-(2-hydroxypropyl)methacrylamide).

[0128] In some embodiments, the degradable functional groups include disulfides, β-thioether esters, amide hydroxymethyl groups, vicinal glycols, peptides, polysaccharides (e.g., alginate, dextran, chitosan, chondroitin), oligosaccharides, o-nitrobenzyl groups, coumarins, acetals, ketals, esters, orthoesters, acid anhydrides, imines, hydrazones, carbonates, azo dyes, carbamates, phosphates, cinnamoyl groups, or benzoyl ethers, or combinations thereof. In some embodiments, the degradable functional groups include disulfides, β-thioether esters, amide hydroxymethyl groups, vicinal glycols, peptides, polysaccharides (e.g., alginate, dextran, chitosan, chondroitin), o-nitrobenzyl groups, coumarins, acetals, ketals, esters, orthoesters, acid anhydrides, imines, hydrazones, carbonates, phosphates, cinnamoyl groups, or benzoyl ethers, or combinations thereof. In some embodiments, the degradable functional groups include disulfides. In some embodiments, the degradable functional group includes a β-thioether ester. In some embodiments, the degradable functional group includes an amide hydroxymethyl group. In some embodiments, the degradable functional group includes a vicinal diol. In some embodiments, the degradable functional group includes a peptide. In some embodiments, the degradable functional group includes one or more polysaccharides.

[0129] In some embodiments, one or more polysaccharides include alginate. In some embodiments, one or more polysaccharides include dextran. In some embodiments, one or more polysaccharides include chitosan. In some embodiments, one or more polysaccharides include chondroitin. In some embodiments, the degradable functional group includes o-nitrobenzyl. In some embodiments, the degradable functional group includes coumarin. In some embodiments, the degradable functional group includes acetal. In some embodiments, the degradable functional group includes ketal. In some embodiments, the degradable functional group includes ester. In some embodiments, the degradable functional group includes orthoester. In some embodiments, the degradable functional group includes acid anhydride. In some embodiments, the degradable functional group includes imine. In some embodiments, the degradable functional group includes hydrazone.

[0130] In some embodiments, the degradable functional group includes carbonates. In some embodiments, the degradable functional group includes phosphates. In some embodiments, the degradable functional group includes cinnamoyl. In some embodiments, the degradable functional group includes benzoyl ether. In some embodiments, the degradable functional group includes carbamates. In some embodiments, the degradable functional group includes azo compounds. In some embodiments, the degradable functional group includes oligosaccharides. In some embodiments, the oligosaccharide includes 2, 3, or 4 monomer units (e.g., 2, 3, or 4 hexoses, pentoses, heptoses, combinations thereof, etc.). In some embodiments, the oligosaccharide includes disaccharides. Non-limiting examples of disaccharides suitable for use as cleavable functional groups include trehalose (which can be cleaved by the enzyme trehalase), maltose (which can be cleaved by the enzyme maltase), sucrose (which can be cleaved by the enzyme sucrase), β-D-mannuronic acid (which can be cleaved by the enzyme enzume alginate), or α-L-guluronic acid (which can be cleaved by the enzyme alginate). In some cases, each arm of the macromonomer includes a PEG group, a triazole, a disaccharide, and a crosslinkable functional group. In specific cases, each arm of the macromonomer, in sequence (with optional intermediate groups), includes a PEG group, a triazole, a disaccharide, and a crosslinkable functional group. For example, each arm of the macromonomer may, in sequence (with optional intermediate groups), include a PEG group, a triazole, a disaccharide, and an acrylate. Alternatively, each arm of the macromonomer may, in sequence (with optional intermediate groups), include a PEG group, a triazole, a disaccharide, and an acrylamide.

[0131] As disclosed herein, oligosaccharide-degradable functional groups endow polymer precursors and the resulting polymer matrix with numerous advantageous properties. Many oligosaccharides are targeted with high specificity by one or more enzymes. In the absence of such enzymes, oligosaccharides can be stable under a wide range of biocompatible conditions, including acidic tumor microenvironments and alkaline washing solutions. Furthermore, because many oligosaccharide-targeting enzymes possess high specificity for individual oligosaccharides or small classes of oligosaccharides, polymer matrices with oligosaccharide-degradable functional groups can be degraded and crosslinked in a highly specific and controlled manner. In many cases, lyases, monooxygenases, hydrolases, or other enzymes targeting oligosaccharide-degradable functional groups may be inactive against other parts of the hydrogel matrix and against other species present in complex biological systems, allowing the polymer matrix to be degraded without affecting other components of the biological system. Therefore, oligosaccharide-degradable functional groups can provide biocompatible polymer matrix cleavage conditions in which cells remain viable. In particular, enzymatic degradation processes in which oligosaccharide-degradable functional groups are enzymatically cleaved generally do not negatively impact cell viability.

[0132] In some embodiments, the crosslinkable functional groups include acrylates, methacrylates, acrylamides, methacrylamides, norbornel, styrene, vinyl ethers, vinylpyrrolidones, vinyl esters, maleimides, allyl groups, alkynes, azides, thiols, olefins, or epoxides or combinations thereof.

[0133] In some embodiments, the crosslinkable functional group includes acrylates. In some embodiments, the crosslinkable functional group includes methacrylates. In some embodiments, the crosslinkable functional group includes acrylamide. In some embodiments, the crosslinkable functional group includes methacrylamide. In some embodiments, the crosslinkable functional group includes norbornyl. In some embodiments, the crosslinkable functional group includes styrene. In some embodiments, the crosslinkable functional group includes vinyl ethers. In some embodiments, the crosslinkable functional group includes vinylpyrrolidone.

[0134] In some embodiments, the crosslinkable functional group includes vinyl esters. In some embodiments, the crosslinkable functional group includes maleimide. In some embodiments, the crosslinkable functional group includes allyl. In some embodiments, the crosslinkable functional group includes alkynes. In some embodiments, the crosslinkable functional group includes azides. In some embodiments, the crosslinkable functional group includes thiols. In some embodiments, the crosslinkable functional group includes olefins. In some embodiments, the crosslinkable functional group includes epoxides.

[0135] In some embodiments, the first stimulus includes an energy source. In some embodiments, the energy source includes a light source. In some embodiments, the wavelength of the light source is between 300 and 500 nm. However, the energy of the light source can be tailored to match the absorbance of the photoinitiator in the polymer precursor formulation. For example, the energy range of the light source can be from about 250 to 700 nm, from about 250 to 300 nm, from about 250 to 400 nm, from about 300 to 450 nm, or from about 400 to 700 nm.

[0136] In some embodiments, the second stimulus comprises an energy source in the presence of a photoinitiator and in the absence of a polymer precursor. In some embodiments, the energy source comprises a light source. In some embodiments, the wavelength of the light source is between 300 and 500 nanometers (nm). In some embodiments, the wavelength of the light source is between 380 nm and 420 nm. However, the energy of the light source can be tailored to match the absorbance of the photoinitiator or photocleavable groups in the polymer matrix. For example, the energy range of the light source can be from about 250 to 700 nm, from about 250 to 300 nm, from about 250 to 400 nm, from about 300 to 450 nm, or from about 400 to 700 nm.

[0137] As used herein, the term "photoinitiator" can refer to a species that generates free radicals based on photoexcitation. In many cases, the photoinitiators included in polymer precursor formulations are either type I photoinitiators (molecules that generate free radicals based on photoexcitation through intramolecular cleavage (e.g., homolytic cleavage)) or type II photoinitiators (molecules that abstract electrons or hydrogen atoms from a co-initiator after photoexcitation). Examples of photoinitiators available in this method include acetophenone, anisole, anthraquinone, anthraquinone-2-sulfonic acid, benzoyl, benzoin, benzophenone, 3,3',4,4'-benzophenone tetracarboxylic dianhydride, 4-benzoylbiphenyl, 2-benzyl-2-(dimethylamino)-4'-morpholinophenylbutanone, dibenzocycloheptenone, 2,2-diethoxyacetophenone, 2-ethylanthraquinone, ferrocene, 2-isopropylthioxanthone, lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), methyl-2-benzoylbenzoate, and thiooxanthen-9-one. In some cases, the photoinitiator in the polymer precursor is a type I photoinitiator. In some cases, the photoinitiator in the polymer precursor is water-soluble. In many cases, the photoinitiator is biocompatible. In specific cases, the photoinitiator in the polymer precursor is lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP).

[0138] In some embodiments, the second stimulus includes the introduction of a degrading agent. In some embodiments, the degrading agent includes dithiothreitol (DTT).

[0139] In some embodiments, the degrading agent includes tris(2-carboxyethyl)phosphine (TCEP). In some embodiments, the degrading agent includes 2-mercaptoethanol (BME). In some embodiments, the degrading agent includes glutathione reductase (GSH). In some embodiments, the degrading agent includes DMEM. In some embodiments, the degrading agent includes RPMI. In some embodiments, the degrading agent includes PBS buffer. In some embodiments, the degrading agent includes sodium (meta)periodate. In some embodiments, the degrading agent includes alginate lyase. In some embodiments, the degrading agent includes dextranase. In some embodiments, the degrading agent includes lysozyme. In some embodiments, the degrading agent includes chitinase. In some embodiments, the degrading agent includes hyaluronidase. In some embodiments, the degrading agent includes chondroitinase.

[0140] In some implementations, the degradation agent includes cellulase.

[0141] In some embodiments, the degradation agent includes an oxidizing agent. The oxidizing agent can be configured to oxidatively cleave one or more degradable functional groups in the polymer matrix, such as one or more vicinal diols. For example, the oxidizing agent may include periodates, such as sodium metaperiodate or sodium orthoperiodate.

[0142] In some embodiments, one or more polymer precursors include a porogen, and increasing the porogen loading results in an increased pore size in one or more polymer matrices. In some embodiments, increasing the porogen concentration increases the diffusion rate by approximately 20%. In some embodiments, the increased pore size increases the diffusion rate of the fluorescently labeled antibody into one or more polymer matrices. In some embodiments, the fluorescently labeled antibody is CD56 PE. In some embodiments, one or more polymer precursors are photopolymerized via photocrosslinking by UV light.

[0143] In some implementations, the method includes chemical or biological materials comprising a first cell in a first internal region and a second cell in a second internal region.

[0144] In some implementations, the method includes a first cell comprising one of a killer cell, an activating cell, and an inhibitory cell, and wherein the first cell interacts with a second cell after degradation in (c).

[0145] In some embodiments, the method includes chemical or biological materials comprising polymer beads containing one or more oligonucleotides arranged thereon.

[0146] In some embodiments, the method includes a chemical or biological material comprising one or more cells or components thereof. In some embodiments, the method further includes one or more polymeric precursors comprising: (a) a macromonomer, (b) a photoinitiator, and (c) a porogen. In some embodiments, the method further includes degradation in (c) caused by cleavage of one or more disulfide bonds in a first gel structure, a second gel structure, or both.

[0147] In some cases, photocuttable 4-arm PEG macromonomers can degrade into cross-linked hydrogel networks containing disulfides. In further cases, the cross-linked hydrogel networks containing disulfides can degrade into networks of soluble fragments.

[0148] In some cases, photocuttable 4-arm PEG macromonomers can be used Figure 21The reaction scheme shown in the figure involves photopolymerization and subsequent degradation into a network of soluble fragments. For simplicity, the crosslinked hydrogel network is depicted in this figure as containing one disulfide in each chain connecting the tertiary carbons. In various embodiments, it is noteworthy that the polymer formed by structure (I) will have two disulfides in each chain connecting the tertiary carbons. In this figure, "PI fragment" represents "photoinitiator fragment". Figure 21 In Chinese, "PI fragment" stands for "photoinitiator fragment".

[0149] In some embodiments, disulfide-based cPEG cages can be degraded using light and photoinitiators. In some cases, degradation allows for spatiotemporal control of cPEG cage degradation, enabling selective retention of cells in a single hydrogel formulation. As used herein, the terms "chamber" and "cage" can refer to a polymer matrix structure that encloses an internal space not filled by a polymer matrix. For example, a chamber or cage can comprise a cylindrical polymer matrix that encloses an internal space not occupied by a polymer matrix, and optionally extends between opposing surfaces of the jetting device such that the internal space is enclosed by the opposing surfaces of the polymer matrix and the jetting device.

[0150] Another aspect of this disclosure provides a kit comprising: (a) a porogen; and (b) a polymer precursor described anywhere in this disclosure.

[0151] The polymer precursor may include monomers selected from structures (I)-(XX). The polymer precursor may include monomers selected from structures (I)-(XIII).

[0152] In some embodiments, the polymer precursor comprises a monomer including an oligomer domain comprising three or more arms, each arm of which comprises a degradable functional group and a crosslinkable functional group. In some cases, the crosslinkable functional group of one of the three or more arms is configured to crosslink with another crosslinkable functional group of another polymer precursor in response to a first stimulus to form a polymeric form of the monomer. In some cases, the degradable functional group is configured to be cleaved in response to a second stimulus, thereby dissolving the polymeric form of the monomer. In some cases, the oligomer domain comprises four or more arms. In some cases, the polymer precursor comprises a monomer selected from structures (I), (II), (IV)-(VI), (XII), (XIII), (XIV)-(XVIII), (XX), and combinations thereof.

[0153] In some cases, degradable functional groups include disulfides, β-thioether esters, amide hydroxymethyl and vicinal diols, vicinal diols, alginate backbones, dextran backbones, chitosan backbones, hyaluronic acid backbones, chondroitin sulfate backbones, or carboxymethyl cellulose backbones, or combinations thereof.

[0154] In some cases, the degradable functional groups include disulfides. After polymer matrix synthesis, the polymer matrix can be cleaved using thiols (such as glutathione or β-mercaptoethanol), disulfides (such as glutathione disulfide), or reducing agents (such as TCEP or dithiothreitol). Such systems offer the advantage of cleavage under mild, biologically compatible conditions, while simultaneously maintaining stability in the absence of disulfide degrading agents.

[0155] In some cases, degradable functional groups include vicinal diols. Compared to hydrolyzable and reductively cleavable functional groups, vicinal diols offer the advantage of enhanced stability, allowing their use under harsh conditions and with cells activated in polymer cleavage. Many cells secrete reactive species capable of degrading reactive functional groups. For example, some neutrophils and macrophages generate the oxidant superoxide extracellularly, while many adherent cells secrete free thiols and regulate extracellular thiol and disulfide concentrations to mediate intercellular signaling. Reactive degradable functional groups such as disulfides and sugars can therefore be unstable in the presence of certain cell types and can exhibit non-negligible background degradation rates in the presence of these cells. The degradable functional groups of vicinal diols often address this issue by being non-reactive to enzymes and molecules secreted by cells. At the same time, vicinal diols are readily cleavable with mild, biocompatible oxidants such as periodate, permanganate, and peroxides in the presence of manganese catalysts. Polymer matrices formed from monomers containing vicinal diols can therefore be formed and degraded rapidly, as disclosed, for example, in Examples 9 and 10.

[0156] In some implementations, the porogen includes poly(ethylene glycol). The molecular weight, degree of crosslinking, and degree of branching of the poly(ethylene glycol) can be selected based on the desired porosity of the polymer matrix to be formed from the polymer precursor.

[0157] In some embodiments, the weight ratio of the porogen to the polymer precursor is between about 1:1 and 1:6. In some embodiments, the weight ratio of the porogen to the polymer precursor is between about 1:2 and 1:5. In some embodiments, the weight ratio of the porogen to the polymer precursor is between about 1:3 and 1:4. In some embodiments, the weight ratio of the porogen to the polymer precursor is about 1:3.5.

[0158] In some embodiments, (a) the porogen and (b) the polymer precursor are contained in a single container. In some embodiments, the kit further includes (c) a photoinitiator contained in another container.

[0159] In some embodiments, the photoinitiator includes one of the following: lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), Irgacure 2959, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide (TPO) nanoparticles, 2,2'-azobis[2-methyl-N-(2-hydroxyethyl)promionamide] (VA-086), BAPO-Oli, BAPO-Ona, eosin Y, riboflavin, and combinations thereof.

[0160] In some embodiments, the oligomeric domain further comprises an amide moiety. In some embodiments, the oligomeric domain further comprises a β-thioether moiety.

[0161] Provided in some embodiments, this document describes a method for processing chemical or biological materials, comprising: (a) inputting (i) the chemical or biological material and (ii) one or more polymer precursors into a jetting device; (b) supplying a stimulus to the jetting device to generate one or more polymer matrices, wherein the one or more polymer matrices comprise: (i) a first gel structure from the one or more polymer precursors within the jetting device, wherein the first gel structure at least partially encapsulates the chemical or biological material, and wherein the first gel structure comprises a first internal region; and (ii) a second gel structure adjacent to the first gel structure, wherein the second gel structure comprises a second internal region; and (c) degrading the first gel structure, the second gel structure, or a portion thereof to generate a third internal region, wherein the third internal region comprises the first internal region of the first gel structure and the second internal region of the second gel structure.

[0162] In some embodiments, the first gel structure and the second gel structure each comprise one or more polymer matrix walls. In some embodiments, the first gel structure and the second gel structure share one or more polymer matrix walls. In some embodiments, a portion of the first gel structure, the second gel structure, or both degraded in (c) includes the shared polymer matrix wall.

[0163] Figures 12A-12D Examples of polymer matrices are shown during cage formation, before degradation, after degradation, and after rinsing. Figure 12A The example shows polymer cage formation. Figure 12B The example shows polymer cage formation of labeled CD56+ cells 1201 prior to degradation. Figure 12C The example shows polymer cage formation of labeled CD56+ cells 1201 after degradation 3 seconds following exposure. Figure 12DThe example shows polymer cage formation of labeled CD56+ cells 1201 after rinsing.

[0164] Figures 12E-12G An example is shown where a portion of the polymer matrix cage degrades to incorporate elements from the polymer matrix. Figure 12F In the example, two adjacent cages are aligned at boundary 1250 to form composite cage 1295. Figure 12F In the example, corner hydrogel markers are used to align the two cages at the boundary 1250 to facilitate the degradation of the partition of the composite cage 1295. Figure 12G The example shows polymer cage formation after degradation following 3 seconds of exposure. (As in...) Figure 12G As shown in the example, boundary 1250 has been substantially degraded, and composite cage 1295 contains a cage with substantially all contents from two adjacent cages.

[0165] In various implementations, processing chemical or biological materials may include sample preparation of biological materials and / or analysis of biological materials.

[0166] In some embodiments, the kit further comprises a polymer precursor that substantially does not contain a free thiol moiety, wherein the free thiol moiety is configured to degrade disulfide bonds. In some embodiments, the individual container substantially does not contain a free thiol moiety, wherein the free thiol moiety is configured to degrade disulfide bonds. In some embodiments, the free thiol concentration is less than 5%, 1%, 0.1%, 0.01%, or 0.001% (e.g., less than 5%, 1%, 0.1%, 0.01%, or 0.001% of disulfide in the macromonomer is reduced to form the free thiol). In some embodiments, the free thiol concentration in the individual container is less than about 10 nM, less than about 5 nM, less than about 1 nM, less than about 500 pM, less than about 100 pM, less than about 50 pM, less than about 10 pM, less than about 5 pM, or less than about 1 pM.

[0167] In one aspect, this document describes a method for processing chemical or biological materials, comprising: (a) introducing (i) the chemical or biological material and (ii) one or more polymeric precursors into a jetting device; (b) supplying a stimulus to the jetting device to generate one or more polymeric matrices from the one or more polymeric precursors within the jetting device, wherein the polymeric matrix of the one or more polymeric matrices at least partially encapsulates the chemical or biological material, and wherein the polymeric matrix is ​​formed within less than 3 seconds after the one or more polymeric precursors come into contact with the stimulus.

[0168] In some embodiments, the polymer matrix is ​​formed within less than 2 seconds after the one or more polymer precursors are contacted with the energy. In some embodiments, the polymer matrix is ​​formed within less than 1 second after the one or more polymer precursors are contacted with the energy. In some embodiments, the polymer matrix is ​​formed within less than 0.5 seconds after the one or more polymer precursors are contacted with the energy.

[0169] In some embodiments, the polymer matrix is ​​formed within less than 0.1 seconds after the one or more polymer precursors come into contact with the energy.

[0170] In one aspect, this document describes a method for treating chemical or biological materials, comprising (a) introducing (i) the chemical or biological material and (ii) one or more polymeric precursors into a jetting device; (b) supplying a stimulus to the jetting device to generate one or more polymeric matrices from the one or more polymeric precursors within the jetting device, wherein the polymeric matrix of the one or more polymeric matrices at least partially encapsulates the chemical or biological material; and (c) performing one or more operations on the chemical or biological material within the polymeric matrix, wherein the one or more operations include a time interval of at least one day, during which no more than 20% of the polymeric matrix degrades.

[0171] In some embodiments, no more than 15% of the polymer matrix degrades during the stated time interval. In some embodiments, no more than 10% of the polymer matrix degrades during the stated time interval. In some embodiments, no more than 5% of the polymer matrix degrades during the stated time interval. In some embodiments, no more than 1% of the polymer matrix degrades during the stated time interval. In some embodiments, the time interval includes at least three days. In some embodiments, the time interval includes at least five days. In some embodiments, the time interval includes at least seven days. In some embodiments, the time interval includes at least fourteen days.

[0172] Figures 1A-1E Illustrative examples are shown in micrographs of polymer matrix compartment formation and encapsulation of one or more biological components within a flow-through chamber over a specified time. In some embodiments, the polymer matrix forms during curing. Figures 1A-1E In the example, it will contain 12.7 mW / cm² at a wavelength of 405 nm. 2 The light source is directed towards a flow cell containing cPEG, PEG (20 kDa), lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), and cells suspended in cell culture medium. The light intensity at the jet device is approximately 100 to 700 mW / cm². 2 .like Figures 1A-1EAs shown in the example, the polymer matrix compartments begin to form after 1 second of exposure. Figure 1A ), and continue to form an exposure lasting up to 5 seconds ( Figures 1B-1E Furthermore, in Figure 1E In the example, the cage is overexposed after 5 seconds. In some cases, longer exposure times are not preferred because the disulfide can be degraded by the photoinitiator after the acrylamide has been converted into a polymer. In various embodiments, the light energy source can emit light with an intensity range from about 10 to 20 mW / cm², preferably 15 to 19 mW / cm² for the first and second stimuli. In a further embodiment, the light source can provide about 50 to 1000 mW / cm² within the jet device. 2 Or approximately 100 to 700 mW / cm 2 The intensity of the first or second stimulus may range from less than 10 seconds, 1-10 seconds, 2-8 seconds, or about 2-4 seconds. The wavelength range of the light energy may be from about 400 nm to about 410 nm, from about 250 to 700 nm, from about 250 to 300 nm, from about 250 to 400 nm, from about 300 to 450 nm, or from about 400 to 700 nm.

[0173] In some cases, the biological component may be localized within a hollow region of the polymer matrix compartment. In other cases, the polymer matrix may be formed on the biological component. In some alternative cases, the polymer matrix may localize more than one biological component. The polymer matrix of the biological compartment may encapsulate one or more biological components. Energy emitted from an energy source may be spatially modulated as described herein to form the polymer matrix. In some cases, the polymer matrix may encapsulate the biological component. In some cases, the biological component may be encapsulated within and / or together with the channel wall of the polymer matrix. In some embodiments, the biological component is released from the cell after exposure to an energy source. In some embodiments, the energy source is UV light for lysing cells. In some embodiments, the energy source is visible light for lysing cells. In some embodiments, UV light is used to activate a photoactivated detergent and lyse cells. In some embodiments, visible light is used to activate a photoactivated detergent and lyse cells.

[0174] In one aspect, this document describes a method for processing chemical or biological materials, comprising: (a) introducing (i) one or more chemical or biological materials and (ii) one or more polymeric precursors into a jetting device; (b) supplying a stimulus to the jetting device to generate one or more polymeric matrices from the one or more polymeric precursors within the jetting device, wherein the one or more polymeric matrices at least partially encapsulate the one or more chemical or biological materials; and (c) performing one or more operations on the one or more chemical or biological materials within the one or more polymeric matrices, wherein the one or more operations include a time interval of at least one day, during which at least 80% of the one or more chemical or biological materials are viable. In some embodiments, at least 85% of the one or more chemical or biological materials are viable during the time interval. In some embodiments, at least 90% of the one or more chemical or biological materials are viable during the time interval. In some embodiments, at least 95% of the one or more chemical or biological materials are viable during the time interval. In some embodiments, at least 99% of the one or more chemical or biological materials are viable during the time interval. In some embodiments, the time interval includes at least three days. In some implementations, the time interval includes at least five days. In some implementations, the time interval includes at least seven days. In some implementations, the time interval includes at least fourteen days.

[0175] Figures 6A-6C Illustrative examples of micrographs showing the encapsulation of polymer matrix compartments and one or more biological components in a flow cell some time after self-cageing. Figures 6A-6C Illustrative examples of micrographs showing the encapsulation of polymer matrix compartments and one or more biological components in a flow-through cell from day 0 to day 2 post-cageing. Figures 6A-6C In the example, K562 cells were minimally exposed to the cPEG precursor solution before cage formation.

[0176] Figures 6D-6F Illustrative examples of micrographs showing the encapsulation of polymer matrix compartments and one or more biological components in a flow cell some time after cage formation. Figures 6D-6F Illustrative examples of micrographs showing the encapsulation of polymer matrix compartments and one or more biological components in a flow-through cell from day 0 to day 2 post-cageing. Figures 6D-6F In the example, K562 cells were exposed to cPEG precursor solution for 3 hours before cage formation. Figures 6A-6F As shown in the example, K562 cells exposed to cPEG precursor solution for 3 hours before caged resulted in similar cell survival and proliferation rates compared to K562 cells with minimal exposure to K562 solution before caged.

[0177] Time interval In some implementations, the time interval includes at least three days. In some implementations, the time interval includes at least five days. In some implementations, the time interval includes at least seven days. In some implementations, the time interval includes at least fourteen days.

[0178] Figures 5A-5D Illustrative examples of micrographs showing the encapsulation of a polymer matrix compartment and one or more biological components in a flow cell over a period of time after formation / curing. Figures 5A-5D Illustrative examples of micrographs showing the encapsulation of polymer matrix compartments and one or more biological components in a flow cell from day 0 to day 14 after formation / curing are displayed. Figure 5A In the example, cPEG cages were formed and Jurkat cells were added on day 0. (As in...) Figures 5A-5D As shown in the example, the formation of cPEG cages allows Jurkat cell proliferation to continue for at least 14 days. Figure 5B In the example, after one day, a small number of Jurkat cells grew into a larger population of Jurkat cells. Furthermore, in Figure 5C In the example, after 5 days, a small number of Jurkat cells grew into a much larger population of Jurkat cells. Furthermore, in... Figure 5D In the example, after 14 days, a small number of Jurkat cells grew into a larger population of Jurkat cells, which essentially comprised the entire volume of the cell cage.

[0179] Figure 5E An illustrative example of a polymer matrix compartment encapsulation of one or more biological components in a flow cell at a certain time after formation / curing is shown. Figure 5E An illustrative example of a polymer matrix compartment encapsulation of one or more biological components in a flow cell is shown in a micrograph at 14 days post-formation / curing. Figure 5E In the example, Jurkat cells were stained with calcein AM to indicate a large number of live cells.

[0180] In one aspect, this document describes a method for processing chemical or biological materials, comprising: (a) feeding (i) one or more chemical or biological materials and (ii) one or more polymer precursors into a jet device; and (b) maintaining jet communication between the one or more chemical or biological materials and the polymer precursors for a duration of at least ten minutes, wherein at least 80% of the one or more chemical or biological materials are viable during the duration.

[0181] In some embodiments, at least 85% of the one or more chemical or biological materials are active during the time interval. In some embodiments, at least 90% of the one or more chemical or biological materials are active during the time interval. In some embodiments, at least 95% of the one or more chemical or biological materials are active during the time interval. In some embodiments, at least 99% of the one or more chemical or biological materials are active during the time interval. In some embodiments, the time interval includes at least 30 minutes. In some embodiments, the time interval includes at least one hour. In some embodiments, the time interval includes at least two hours. In some embodiments, the time interval includes at least three hours. In some embodiments, the time interval includes at least one day. In some embodiments, the method further includes supplying a stimulus to the jetting device to generate one or more polymer matrices from the one or more polymer precursors within the jetting device, wherein the one or more polymer matrices at least partially encapsulate the one or more chemical or biological materials. In some embodiments, the stimulus includes light. In some embodiments, the one or more polymer matrices comprise a polymeric form of a cPEG monomer.

[0182] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (I): .

[0183] In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10.

[0184] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (II): (II).

[0185] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10.

[0186] In some embodiments, n is between about 0 and about 10. In some embodiments, one or more polymer precursors comprise a solution of at least 40 µL.

[0187] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (III): (III).

[0188] In some cases, n and m are independently integers selected from 0 to 500, where at least one of n and m is at least 1. In some cases, n and m are independently integers selected from 0 to 100, where at least one of n and m is at least 1. In some cases, n and m are independently integers selected from 1 to 100. In some cases, n and m are independently integers selected from 5 to 100. In some cases, n and m are independently integers selected from 5 to 50. In some cases, n and m are independently integers selected from 20 to 50. In some cases, n and m are independently integers selected from 0 to 10, where at least one of n and m is at least 1.

[0189] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (IV): .

[0190] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10. In some cases, each instance of m is an integer independently selected from 0 to 500. In some cases, each instance of m is an integer independently selected from 0 to 100. In some cases, each instance of m is an integer independently selected from 1 to 100. In some cases, each instance of m is an integer independently selected from 5 to 100. In some cases, each instance of m is an integer independently selected from 5 to 50. In some cases, each instance of m is independently an integer selected from 20 to 50. In other cases, each instance of m is independently an integer selected from 0 to 10.

[0191] In some embodiments, the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising a structure (V): .

[0192] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10.

[0193] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (VI): .

[0194] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10.

[0195] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (VII): (VII)

[0196] In some cases, structure (VII) includes between 1 and 500, 1 to 100, 5 to 100, 5 to 50, 20 to 50, or 1 to 10 monolithic units.

[0197] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (VIII): .

[0198] In some cases, n, m, and p are independently integers selected from 0 to 500, where at least one of n, m, and p is at least 1. In some cases, n, m, and p are independently integers selected from 0 to 100, where at least one of n, m, and p is at least 1. In some cases, n and m are independently integers selected from 1 to 100. In some cases, n, m, and p are independently integers selected from 5 to 100. In some cases, n, m, and p are independently integers selected from 5 to 50. In some cases, n, m, and p are independently integers selected from 20 to 50. In some cases, n, m, and p are independently integers selected from 0 to 10, where at least one of n, m, and p is at least 1.

[0199] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising the structure (IX): (IX).

[0200] In some cases, n is an integer selected from 1 to 500. In some cases, n is an integer selected from 1 to 100. In some cases, n is an integer selected from 5 to 100. In some cases, n is an integer selected from 5 to 50. In some cases, n is an integer selected from 20 to 50. In some cases, n is an integer selected from 1 to 10.

[0201] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (X): (X).

[0202] In some cases, n and m are independently integers selected from 0 to 500, where at least one of n and m is at least 1. In some cases, n and m are independently integers selected from 0 to 100, where at least one of n and m is at least 1. In some cases, n and m are independently integers selected from 1 to 100. In some cases, n and m are independently integers selected from 5 to 100. In some cases, n and m are independently integers selected from 5 to 50. In some cases, n and m are independently integers selected from 20 to 50. In some cases, n and m are independently integers selected from 0 to 10, where at least one of n and m is at least 1.

[0203] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (XI): .

[0204] In some cases, the structure (XI) includes between 1 and 500, 1 to 100, 5 to 100, 5 to 50, 20 to 50, or 1 to 10 monosaccharide units.

[0205] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (XII): .

[0206] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10.

[0207] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising structure (XIII): .

[0208] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10.

[0209] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising a structure (XIV): Where R 1 Yes: i) C1 alkyl group, ii) C2-C 18 Linear or branched saturated alkyl groups, or (iii) C3-C8 cyclic saturated alkyl groups substituted by 0-4 independently selected C1-C3 alkyl groups; wherein R 1 quilt Replace q instances of R; 1 The remaining substituents are hydrogen, q is an integer from 2 to 32, and each instance of n is an integer selected from 0 to 500. In some cases, q is an integer from 2 to 10. In some cases, q is an integer from 2 to 6. In some cases, q is an integer from 3 to 5. In some cases, R 1 It is a C1 alkyl group or a C2-C6 saturated linear or branched alkyl group, and q is an integer from 2 to 14. In some cases, R 1 It is a C1-C2 alkyl group, and q is an integer from 2 to 6. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50.

[0210] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising a structure (XV): , Where R 2 Yes: i) C1 alkyl group, ii) C2-C 18 Linear or branched saturated alkyl groups, or iii) C3-C8 cyclic saturated alkyl groups substituted by 0-4 independently selected C1-C3 alkyl groups; Where R 2 quilt Replace w instances of R 2 The remaining substituents are hydrogen, w is an integer from 2 to 32, each instance of n is an integer independently selected from 0 to 500, and each instance of m is an integer independently selected from 0 to 500. In some cases, w is an integer from 2 to 10. In some cases, w is an integer from 2 to 6. In some cases, w is an integer from 3 to 5. In some cases, R 2 It is a C1 alkyl group or a C2-C6 saturated linear or branched alkyl group, and w is an integer from 2 to 14. In some cases, R 2 It is a C1-C2 alkyl group, and w is an integer from 2 to 6. In some cases, n is an integer selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer selected from 5 to 50. In some cases, each instance of n is an integer selected from 20 to 50. In some cases, each instance of m is an integer independently selected from 0 to 500. In some cases, each instance of m is an integer independently selected from 0 to 100. In some cases, each instance of m is an integer independently selected from 20 to 50.

[0211] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising a structure (XVI): , Where R 3 Yes: i) C1 alkyl group, ii) C2-C 18 Linear or branched saturated alkyl groups, or iii) C3-C8 cyclic saturated alkyl groups substituted by 0-4 independently selected C1-C3 alkyl groups; Where R 3 quilt Replace x instances of R 3 The remaining substituents are hydrogen, x is an integer from 2 to 32, and each instance of n is an integer independently selected from 0 to 500. In some cases, x is an integer from 2 to 10. In some cases, x is an integer from 2 to 6. In some cases, x is an integer from 3 to 5. In some cases, R 3 It is a C1 alkyl group or a C2-C6 saturated linear or branched alkyl group, and x is an integer from 2 to 14. In some cases, R 3It is a C1-C2 alkyl group, and x is an integer from 2 to 6. In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50.

[0212] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising the structure (XVII): .

[0213] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10.

[0214] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising a structure (XVIII): .

[0215] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10. In some cases, each instance of m is an integer independently selected from 0 to 500. In some cases, each instance of m is an integer independently selected from 0 to 100. In some cases, each instance of m is an integer independently selected from 1 to 100. In some cases, each instance of m is an integer independently selected from 5 to 100. In some cases, each instance of m is an integer independently selected from 5 to 50. In some cases, each instance of m is independently an integer selected from 20 to 50. In other cases, each instance of m is independently an integer selected from 0 to 10.

[0216] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising a structure (XIX): .

[0217] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10. In some cases, each instance of m is an integer independently selected from 0 to 500. In some cases, each instance of m is an integer independently selected from 0 to 100. In some cases, each instance of m is an integer independently selected from 1 to 100. In some cases, each instance of m is an integer independently selected from 5 to 100. In some cases, each instance of m is an integer independently selected from 5 to 50. In some cases, each instance of m is independently an integer selected from 20 to 50. In other cases, each instance of m is independently an integer selected from 0 to 10.

[0218] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer comprising a structure (XX): .

[0219] In some cases, each instance of n is an integer independently selected from 0 to 500. In some cases, each instance of n is an integer independently selected from 0 to 100. In some cases, each instance of n is an integer independently selected from 1 to 100. In some cases, each instance of n is an integer independently selected from 5 to 100. In some cases, each instance of n is an integer independently selected from 5 to 50. In some cases, each instance of n is an integer independently selected from 20 to 50. In some cases, each instance of n is an integer independently selected from 0 to 10. In some cases, each instance of m is an integer independently selected from 0 to 500. In some cases, each instance of m is an integer independently selected from 0 to 100. In some cases, each instance of m is an integer independently selected from 1 to 100. In some cases, each instance of m is an integer independently selected from 5 to 100. In some cases, each instance of m is an integer independently selected from 5 to 50. In some cases, each instance of m is independently an integer selected from 20 to 50. In other cases, each instance of m is independently an integer selected from 0 to 10.

[0220] In some embodiments, one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising: an oligomer domain having three or more arms, each arm of said oligomer domain comprising a degradable functional group and a crosslinkable functional group, wherein the crosslinkable functional group of one of the three or more arms is configured to crosslink with another crosslinkable functional group of another polymer precursor in response to a first stimulus to obtain the polymeric form of the monomer, and wherein the degradable functional group is configured to be cleaved in response to a second stimulus to dissolve the polymeric form of the monomer.

[0221] In some implementations, the oligodomain contains four or more arms.

[0222] In some embodiments, one or more polymer matrices comprise degradable functional groups. In some embodiments, the degradable functional groups include disulfides, β-thioether esters, amide hydroxymethyl and vicinal diols, vicinal diols, alginate backbones, dextran backbones, chitosan backbones, hyaluronic acid backbones, chondroitin sulfate backbones, or carboxymethyl cellulose backbones, or combinations thereof. In some embodiments, one or more polymer matrices comprise a hydrogel. In some embodiments, the hydrogel comprises cPEG, cSEL-BTEEC, cSEL-DHEBA, cSEL-diol, cSEL-alginate, cSEL-dextran, cSEL-chitosan, cSEL-hyaluronic acid, cSEL-chondroitin sulfate, or cSEL-cellulose, or combinations thereof. In some embodiments, one or more polymer matrices comprise a degradation agent. In some embodiments, the degradation agents include DTT, TCEP, BME, GSH, DMEM, RPMI, PBS buffer, DMEM, RPMI, PBS buffer, (meta)periodate, alginate lyase (enzyme), dextranase, lysozyme and chitinase, hyaluronidase, chondroitinase or cellulase or combinations thereof.

[0223] In some embodiments, one or more polymer matrices comprise at least one β-thioether ester. In some embodiments, one or more polymer matrices comprise a PEG moiety containing a β-thioether ester. In some embodiments, the β-thioether ester is formed by reacting an acrylate with a thiol. In some embodiments, one or more polymer matrices are degraded overnight in the presence of a commonly used cell culture medium, thereby providing a biocompatible degradation pathway for selective retention experiments. In some embodiments, one or more polymer matrices comprise a Michael donor. In some embodiments, the Michael donor is a PEG-thiol. In some embodiments, one or more polymer matrices comprise a cSEL β-thioether ester having one β-thioether ester per arm. In some embodiments, one or more polymer matrices comprise any material formed of PEG containing a Michael acceptor chain. In some embodiments, the Michael acceptor chain comprises PEG-acrylamide, PEG-vinyl sulfone, PEG-maleimide, or PEG-carbonylacrylic acid, or any combination thereof.

[0224] In some embodiments, one or more polymer matrices are degraded by cleavage of disulfide bonds. In some embodiments, disulfide bonds are cleaved by one or more reducing agents. In some embodiments, one or more reducing agents include DTT, TCEP, BME, or GSH, or any combination thereof. In some embodiments, one or more polymer matrices comprise one or more arms, each containing one or more amides. In some embodiments, one or more polymer matrices are degraded by oxidative cleavage of vicinal diols by sodium periodate. In some embodiments, the method further includes: (a) using a caged workflow, (b) performing selective cell retention when used with cPEG or other hydrogels having an orthogonal degradation mechanism, or (c) performing long-term cell incubation using a reducing agent that does not affect cage stability in the cell culture medium. As an illustrative example, a “caged workflow” may include loading a polymer precursor and one or more cells into a jet device, projecting light into the jet device using a spatial energy modulation element (e.g., a digital micromirror device), such that the projected light causes crosslinking of the polymer precursor to at least partially form one or more cages or chambers enclosing at least a subset of one or more cells. One or more cages or chambers may be formed at locations identified by the detector, for example, at locations at least around a subset of one or more cells.

[0225] In some embodiments, one or more polymer matrices comprise vicinal diol functionality that can be cleaved via oxidation of one or more hydroxyl groups. In some embodiments, one or more polymer matrices are used to formulate stable hydrogels. In some embodiments, one or more polymer matrices comprise a polymeric form of a photocleavable 4-arm PEG-monomer. In some embodiments, one or more polymer matrices are photodegraded via a vicinal nitrobenzyl moiety. In some embodiments, one or more polymer matrices comprise a polymeric form of a coumarin-based photodegradable monomer. In some embodiments, one or more polymer matrices comprise a 4-arm PEG-acrylamide comprising one or more disulfides. In some embodiments, one or more polymer matrices comprise one or more cage disulfide bonds in the formation of hydrogel cages, wherein the hydrogel cages are degraded using light and a photoinitiator. In some embodiments, one or more polymer matrices enable hydrogel formation, and wherein the hydrogel enables spatiotemporal control of hydrogel cage degradation, thus enabling selective retention of cells with a single hydrogel formulation. In some embodiments, one or more polymer matrices enable hydrogel formation, and wherein the use of light and a photoinitiator as a reagent enables the hydrogel to selectively degrade cages and release their contents.

[0226] In some embodiments, one or more polymer precursors enable the formation of hydrogels up to 2 mm thick, and wherein the hydrogel can be formed within 120 s with low light intensity (e.g., 10 mW / cm² from a light source at 265 nm). 2 Or approximately 50 to 1000 mW / cm at the jet device 2 Or approximately 100 to 70 mW / cm 2 ) is degraded.

[0227] In some implementations, when exposed to light, photogenerated free radicals initiate multiple fracture and disulfide exchange reactions, thereby allowing and promoting photodeformation, photowelding, and photodegradation of one or more polymer matrices.

[0228] In some implementations, one or more polymer precursors enable the formation of hydrogels, wherein the hydrogels exhibit chemical or physical changes in response to external stimuli.

[0229] In some embodiments, one or more polymer matrices include photoinstantaneous nitrobenzyl esters that cleave upon photon absorption, thereby allowing the user to exogenously control the degradation of one or more polymer matrices.

[0230] In some embodiments, one or more polymer precursors enable hydrogel formation, and the method further includes controlling the network degradation of the hydrogel by adjusting the concentration of a photoinitiator infused into the hydrogel.

[0231] In some embodiments, one or more polymer precursors contain thiol groups that are quenched after the formation of one or more polymer matrices.

[0232] In some implementations, higher concentrations of bisacrylamide in one or more polymer matrices result in larger pore sizes.

[0233] In some embodiments, the degradation of one or more polymer matrices depends on the type of encapsulated cells, the number of cells, or the fraction of disulfide moieties present in one or more polymer matrices. In some embodiments, the grafting efficiency of one or more polymer matrices is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.

[0234] Figure 2A The effect of buffer concentration in the polymer matrix precursor solution on the degradation rate of the polymer matrix is ​​shown. Figure 2A The effect of buffer concentration in the cPEG precursor solution on the degradation rate of the hydrogel is shown. Figure 2AIn the example, a cell-compatible degradation agent was used. In some implementations, the degradation agent includes glutathione (“GSH”). Further, Figure 2A The example illustrates the lysis of a 10% cPEG gel by 1 mM and 10 mM GSH in Tris pH 8.6. In some embodiments, increasing the concentration of the degrading agent (e.g., GSH) reduces the gel lysis time. Figure 2A As shown in the example, increasing the concentration of GSH from 1 mM to 10 mM in the working solution (e.g., the precursor solution) reduced the gel lysis time (min) of the 2s cage from 18 minutes to 2 minutes. Figure 2A The example further shows that increasing the concentration of GSH from 1 mM to 10 mM in the working solution (e.g., the precursor solution) reduced the gel lysis time (min) of the 4s cage from 24 min to 2.5 min.

[0235] Figure 2B The degradation of the polymer matrix by the degradation reagent is shown in buffers at different pH values. Figure 2B Examples show the lysis of a 10% cPEG gel by 10 mM GSH and 10 mM GSH in Tris at pH 8.0 and 8.6. In some embodiments, reducing the pH of the working solution (e.g., the precursor solution) increases the gel lysis time. Figure 2B As shown in the example, reducing the pH of the working solution (e.g., the precursor solution) from 8.6 to 8.0 increases the gel lysis time (min) of the 2s cage from 2 minutes to 4 minutes. Figure 2B The example further shows that reducing the pH of the working solution (e.g., the precursor solution) from 8.6 to 8.0 increases the gel lysis time (min) of the 4s cage from 2.5 min to 5 min.

[0236] In some embodiments, one or more polymer precursors are in a solution with a pH of at least 8.0.

[0237] Figure 3 The degradation of the polymer matrix by the degradation reagent is shown in buffers at different pH values. Figure 3 The example illustrates the lysis of a 10% cPEG gel by 10 mM GSH in pH-adjusted RPMI 1640 cell culture medium / PBS. In some embodiments, increasing the pH of the working solution (e.g., the precursor solution) reduces the gel lysis time. Figure 3 As shown in the example, increasing the pH of the working solution (e.g., the precursor solution) from 6.8 to 8.1 in the working solution reduces the gel lysis time (min) of the 2s cage from 80 min to 4 min. Figure 3The example further shows that increasing the pH of the working solution (e.g., the precursor solution) from 7.5 to 8.1 reduced the gel lysis time (min) of the 4s cage from 17 minutes to 5 minutes.

[0238] Figure 4A This is an illustrative graph comparing gel lysis time in minutes on the Y-axis with the pH of the working solution on the X-axis for a representative polymer matrix. Figure 4A This is an illustrative graph comparing the gel lysis time in minutes on the Y-axis with the pH of the working solution on the X-axis (2S cage). Figure 4A The example shows the lysis of a 10% cPEG gel by 10 mM GSH in pH-adjusted RPMI 1640 cell culture medium / PBS. In some embodiments, increasing the pH of the working solution (e.g., the precursor solution) reduces the gel lysis time. Furthermore, as... Figure 4A As shown in the examples, the optimal working solution pH range for the 2S cage can include between about pH 7.5 and about pH 8.1. In some embodiments, the gel lysis time of the 2S cage between about pH 7.5 and about pH 8.1 can include up to about 15 minutes.

[0239] Figure 4B This is an illustrative graph comparing gel lysis time in minutes on the Y-axis with the pH of the working solution on the X-axis for a representative polymer matrix. Figure 4B This is an illustrative graph comparing the gel lysis time in minutes on the Y-axis with the pH of the working solution on the X-axis using a 4S cage. Figure 4B The example shows the lysis of a 10% cPEG gel by 10 mM GSH in pH-adjusted RPMI 1640 cell culture medium / PBS. In some embodiments, increasing the pH of the working solution (e.g., the precursor solution) reduces the gel lysis time. Furthermore, as... Figure 4B As shown in the examples, the optimal working solution pH range for the 4S cage can include between about pH 7.8 and about pH 8.1. In some embodiments, the gel lysis time of the 4S cage between about pH 7.8 and about pH 8.1 can include up to about 10 minutes.

[0240] Polymer precursors may include porogens. As used herein, the term "porogen" can refer to a species that modulates the porosity of a polymer matrix. Porogens may be dispersed with the reactants prior to the polymerization process that forms the polymer matrix. After polymerization, porogens typically diffuse out of the polymer matrix, leaving pores in the areas they occupy. The size, concentration, hydrophobicity, and hydrophilicity of the porogen can therefore affect the pore density and pore size in the polymer matrix. Examples of porogens consistent with this disclosure include particles (e.g., polymers, ceramics, metals, metal oxides, or hydrogel particles), polymers (such as polyethylene glycol and alginate), and vesicles (such as liposomes or micelles).

[0241] Figure 7 This is an illustrative graph comparing the percentage of background intensity on the Y-axis with the diffusion time (min) of one or more biological components into the polymer matrix on the X-axis. Figure 7 This is an illustrative graph comparing the percentage of background intensity on the Y-axis with the diffusion time (min) of one or more biological components into the hydrogel cages on the X-axis. Figure 7 In the examples, biological components may include CD56PE (dye-labeled antibody). Figure 7 As shown, a working solution 701 loaded with a first porogen is compared with a working solution 702 loaded with a second porogen. Figure 7 In the example, the porogen-loaded working solution 702 contains 50% more porogen by volume than the porogen-loaded working solution 702. For example... Figure 7 As shown, increasing the porogen loading in the working solution increases the diffusion rate of CD56PE into the cage. Furthermore, in some embodiments, increasing the porogen loading can increase the pore size of the cage.

[0242] Figures 8A-8D Illustrative example of a polymer-matrix compartment encapsulation of one or more biological components in a flow cell. Figures 8A-8D An illustrative example of a polymer-matrix compartment encapsulation of one or more biological components in a flow cell is shown in a photographic print. Figure 8A In the example, the precursor solution contains a nominal porogen (e.g., a working solution 701 loaded with a first porogen) and is used to generate hydrogel cages. Figure 8B In the example, the precursor solution contains more than 50% porogen (e.g., a working solution 702 loaded with a second porogen) and is used to generate hydrogel cages, the precursor solution being in combination with... Figure 8A The ones used to form hydrogels are similar. Figure 8CThe example is an illustrative example of multiple hydrogel cages after 1 minute of adding CD56PE (dye-labeled antibody) at a concentration of about 30 µL / mL to the outside of the hydrogel cage, wherein the hydrogel cages are made with a precursor solution containing a nominal porogen (e.g., working solution 701 loaded with a first porogen). Figure 8D The example is an illustrative example of multiple hydrogel cages 10 minutes after CD56PE was added to the outside of the hydrogel cage at a concentration of approximately 30 µL / mL, wherein the precursor solution contains a nominal porogen (e.g., working solution 701 loaded with a first porogen). Figure 8C As shown, the interior of the cage is dark, indicating almost no fluorescence from CD56 PE within the cage, while the outer gaps are bright, indicating the presence of CD56 PE. Figure 8D As shown, the inner portion of the cage is bright, indicating fluorescence from CD56 PE within the cage's interior. After 10 minutes, measurable amounts of CD56 PE diffuse through the pores of the hydrogel cage. Therefore, CD56 PE fluorescence was observed in the interstitial spaces outside the cage and inside the cage, as shown in... Figure 8D The area is shown against a bright background. Notably, dark rings form on the walls of the hydrogel chambers, indicating low levels of CD56 PE.

[0243] Figures 9A-9B An illustrative example of the volumetric composition of a precursor solution for forming a polymer matrix compartment configured to encapsulate one or more biological components in a flow cell is shown. Figures 9A-9B An illustrative example of the volumetric composition of a precursor solution for forming a polymer matrix compartment configured to encapsulate one or more biological components in a flow cell is shown.

[0244] exist Figure 9A The example shows the composition of the first porogen-loaded working solution 701. In some embodiments, the first porogen-loaded working solution 701 comprises about 21 µL of 20% w / v cPEG stock solution, about 6 µL of porogen stock solution (25% w / v), and about 9 µL of phosphate-buffered saline (PBS). In some embodiments, the first porogen-loaded working solution 701 comprises a total volume of about 36 µL. In some embodiments, the first porogen-loaded working solution 701 comprises about 58.3% cPEG stock solution by volume, about 16.7% porogen stock solution (25% w / v) by volume, and about 25% PBS by volume.

[0245] exist Figure 9AThe example shows the composition of the second porogen-loaded working solution 702. In some embodiments, the second porogen-loaded working solution 702 comprises about 21 µL of 20% w / v cPEG stock solution, about 9 µL of porogen stock solution (25% w / v), and about 6 µL of PBS. In some embodiments, the first porogen-loaded working solution 701 comprises a total volume of about 36 µL. In some embodiments, the second porogen-loaded working solution 702 comprises about 58.3% cPEG stock solution by volume, about 25% porogen stock solution (25% w / v) by volume, and about 16.7% PBS by volume.

[0246] Figure 9B An illustrative example of the volumetric composition of a precursor solution for forming a polymer matrix compartment configured to encapsulate one or more biological components in a flow cell is shown. Figure 9B In the example, precursor solution 901 contains approximately 30% by volume of working solution 701 loaded with a first porogen, approximately 20% by volume of working solution 702 loaded with a second porogen, and approximately 50% by volume of PBS. Figure 9B In the example, precursor solution 1001 contains approximately 36 µL of working solution 701 loaded with a first porogen, approximately 24 µL of working solution 702 loaded with a second porogen, and approximately 60 µL of PBS. Figure 9B In the example, precursor solution 1001 comprises a total volume of approximately 120 µL.

[0247] cPEG concentration Figure 10A This is an illustrative graph comparing the % background intensity on the Y-axis with the diffusion time (min) of one or more biological components into the polymer matrix on the X-axis. Figure 10A This is an illustrative graph comparing the % background intensity on the Y-axis with the diffusion time (min) of one or more biological components into the hydrogel cage on the X-axis. Figure 10A In the example, multiple cPEG-loaded solutions are plotted, comparing the % background intensity on the Y-axis with the diffusion time (min) of one or more biological components into the hydrogel cage on the X-axis. In some embodiments, the multiple cPEG-loaded solutions include a second cPEG-loaded solution 1102, a third cPEG-loaded solution 1103, and a fourth cPEG-loaded solution 1104.

[0248] Figure 10B It is an illustrative graph that compares the % background intensity on the Y-axis at fixed time intervals with the concentration of the polymer precursor stock solution on the X-axis, according to some implementation schemes. Figure 10BThis is an illustrative graph comparing the % background intensity on the Y-axis at 10 min with the concentration of the cPEG stock solution on the X-axis. (See figure below.) Figure 10B As shown in Figure 1050, increasing the concentration of cPEG loading reduced the diffusion of biological components into the hydrogel cages by 10 min.

[0249] Figure 11 Illustrative examples of precursor solution compositions by volume for forming polymer matrix compartments configured to encapsulate one or more biological components in a flow cell, according to some embodiments, are shown.

[0250] More specifically, Figure 11 Illustrative examples of four polymer precursor solution compositions by volume are shown. These polymer precursor solution compositions can be used to form polymer matrix compartments to encapsulate one or more biological components (e.g., prokaryotic or eukaryotic cells) in a flow cell. Figure 11 The examples show four cPEG-loaded solutions: a first cPEG-loaded solution 1101, a second cPEG-loaded solution 1102, a third cPEG-loaded solution 1103, and a fourth cPEG-loaded solution 1104. Each solution contains a small number of beads, which serve as a reference structure during imaging.

[0251] In some embodiments, the second cPEG-loaded solution 1102 contains a nominal concentration of cPEG stock solution (20% w / v). In a further embodiment, the first cPEG-loaded solution 1102 contains a cPEG stock solution (20% w / v) at a concentration approximately 0.75 times that of the second cPEG-loaded solution 1102. In a further embodiment, the third cPEG-loaded solution 1103 contains a cPEG stock solution (20% w / v) at a concentration approximately 1.1 times that of the second cPEG-loaded solution 1102. In a further embodiment, the fourth cPEG-loaded solution 1104 contains a cPEG stock solution (20% w / v) at a concentration approximately 1.2 times that of the second cPEG-loaded solution 1102.

[0252] exist Figure 11 In one example, the first cPEG-loaded working solution 1101 comprises a total volume of approximately 120.6 µL. In some embodiments, the first cPEG-loaded working solution 1101 comprises approximately 14.6 µL of cPEG stock solution (20% w / v), approximately 6.00 µL of porogen stock solution (e.g., 25% w / v), approximately 12.0 µL of LAP solution (3 mg / mL), approximately 87.4 µL of PBS buffer, and approximately 0.6 µL of beads.

[0253] exist Figure 11In one example, the second cPEG-loaded working solution 1103 comprises a total volume of approximately 120.6 µL. In some embodiments, the second cPEG-loaded working solution 1102 comprises approximately 19.5 µL of cPEG stock solution (20% w / v), approximately 6.00 µL of porogen stock solution (e.g., 25% w / v), approximately 12.0 µL of LAP solution (3 mg / mL), approximately 82.65 µL of PBS buffer, and approximately 0.6 µL of beads.

[0254] exist Figure 11 In one example, the third cPEG-loaded working solution 1104 comprises a total volume of approximately 120.7 µL. In some embodiments, the third cPEG-loaded working solution 1103 comprises approximately 21.5 µL of cPEG stock solution (20% w / v), approximately 6.00 µL of porogen stock solution (e.g., 25% w / v), approximately 12.0 µL of LAP solution (3 mg / mL), approximately 80.6 µL of PBS buffer, and approximately 0.6 µL of beads.

[0255] exist Figure 11 In one example, the fourth cPEG-loaded working solution 1104 comprises a total volume of approximately 120.6 µL. In some embodiments, the fourth cPEG-loaded working solution 1104 comprises approximately 23.4 µL of cPEG stock solution (20% w / v), approximately 6.00 µL of porogen stock solution (e.g., 25% w / v), approximately 12.0 µL of LAP solution (3 mg / mL), approximately 78.6 µL of PBS buffer, and approximately 0.6 µL of beads.

[0256] Another aspect of this disclosure provides a flow cell for sequencing genes, comprising: multiple cells, multiple nanopores, and one or more polymer precursors; wherein a polymer matrix is ​​formed by the one or more polymer precursors within less than 10 seconds after contact with energy. In some embodiments, the jet device is a microjet device or a nanojet device. In some embodiments, the jet device includes a sequencing flow cell. In some embodiments, the jet device is used for nucleic acid sequencing. In some embodiments, the biological components include cells, nucleic acids, microbiomes, proteins, cellular assemblages, spatially connected biological components, or metabolites. In some embodiments, the cells are animal cells (e.g., human cells), plant cells, fungal cells, bacterial cells, tumor spheroids, or combinations thereof. In some embodiments, the nucleic acids are DNA of 100 base pairs or more or RNA of 50 base pairs or more.

[0257] In some embodiments, the jet device may be a flow cell. For example, the jet device may be used for sequencing (e.g., DNA or RNA sequencing). In some embodiments, the jet device may be a microjet device. In some embodiments, the jet device may be a nanojet device.

[0258] This disclosure provides computer systems that are programmed to implement the methods of this disclosure. Figure 13 The computer system 1501 is displayed and can be programmed or otherwise configured to perform the methods described herein. The computer system 1501 can be modulated to various aspects of this disclosure, such as identifying biological components, detecting barcodes, generating spatial modulation elements (e.g., masks), providing energy from an energy source, or using sensors to detect or measure local parameters. The detector may be a camera (e.g., a fluorescence camera). The computer system 1501 may be an electronic device belonging to a user or a computer system that may be located remotely relative to the electronic device. The electronic device may be a mobile electronic device.

[0259] Computer system 1501 includes a central processing unit (CPU, also referred to herein as a “processor” and “computer processor”) 1505, which may be a single-core or multi-core processor, or multiple processors for parallel processing. Computer system 1501 also includes memory or storage location 1510 (e.g., random access memory, read-only memory, flash memory), electronic storage unit 1515 (e.g., hard disk), communication interface 1520 for communicating with one or more other systems (e.g., network adapter), and peripheral devices 1525, such as cache, other memory, data storage, and / or electronic display adapter. Memory 1510, storage unit 1515, interface 1520, and peripheral devices 1525 communicate with CPU 1505 via a communication bus (solid line) (such as a motherboard). Storage unit 1515 may be a data storage unit (or data repository) for storing data. Computer system 1501 can be operatively connected to computer network (“network”) 1530 via communication interface 1520. Network 1530 may be the Internet, an internet and / or an extranet, or an intranet and / or extranet that can communicate with the Internet. In some cases, network 1530 may be a telecommunications and / or data network. Network 1530 may include one or more computer servers that can enable distributed computing, such as cloud computing. In some cases, with the aid of computer system 1501, network 1530 can enable a peer-to-peer network, which allows devices connected to computer system 1501 to act as clients or servers.

[0260] CPU 1505 can execute machine-readable instruction sequences, which can be embodied in a program or software. Instructions can be stored in a memory location (such as memory 1510). Instructions can be directed to CPU 1505, which can then be programmed or otherwise configured to implement the methods of this disclosure. Examples of operations performed by CPU 1505 can include fetching, decoding, executing, and writing back.

[0261] CPU 1505 may be part of a circuit, such as an integrated circuit. One or more other components of system 1501 may be included in the circuit. In some cases, the circuit may be an application-specific integrated circuit (ASIC).

[0262] Storage unit 1515 can store files, such as drivers, libraries, and saved programs. Storage unit 1515 can store user data, such as user preferences and user programs. In some cases, computer system 1501 may include one or more additional data storage units external to computer system 1501, such as those located on a remote server that can communicate with computer system 1501 via an intranet or the Internet.

[0263] Computer system 1501 can communicate with one or more remote computer systems via network 1530. For example, computer system 1501 can communicate with a user's remote computer system (e.g., a laptop computer, personal computer, tablet computer, or mobile phone). Examples of remote computer systems include personal computers (e.g., portable PCs), slate / tablet PCs (e.g., Apple® iPad, Samsung® Galaxy Tab), telephones, smartphones (e.g., Apple® iPhone, Android-enabled devices, Blackberry®), or personal digital assistants. Users can access computer system 1501 via network 1530.

[0264] The methods described herein can be implemented by machine-executable code (e.g., a computer processor) stored at an electronic storage location in computer system 1501 (e.g., stored in memory 1510 or electronic storage unit 1515). The machine-executable or machine-readable code can be provided in software form. During use, the code can be executed by processor 1505. In some cases, the code can be retrieved from storage unit 1515 and stored in memory 1510 for access by processor 1505. In some cases, electronic storage unit 1515 can be excluded, and machine-executable instructions are stored in memory 1510.

[0265] The code can be pre-compiled and configured for use with a machine having a processor suitable for executing the code, or it can be compiled at runtime. The code can be provided in a programming language, which can be selected to enable the code to be executed either pre-compiled or compiled.

[0266] Aspects of the systems and methods provided herein, such as computer system 1501, can be implemented in a program. Various aspects of the technology can be considered "products" or "articles of art," typically in the form of machine (or processor) executable code and / or associated data, which can be carried or contained in a type of machine-readable medium. Machine-executable code can be stored on electronic storage units such as memory (e.g., read-only memory, random access memory, flash memory) or hard disks. "Storage" type media can include any or all tangible memory or associated modules of computers, processors, etc., such as various semiconductor memories, tape drives, disk drives, etc., which can provide non-transitory storage for software programming at any time. All or part of the software can sometimes be communicated via the Internet or various other telecommunications networks. For example, such communication can enable the loading of software from one computer or processor into another computer or processor, e.g., from a management server or host computer into a computer platform for an application server. Therefore, another type of medium that can carry software elements includes optical, electrical, and electromagnetic waves, such as those used on physical interfaces between local devices via wired and optical terrestrial networks and various air links. Physical elements carrying such waves, such as wired or wireless links, optical links, etc., can also be considered as media carrying software. As used herein, unless limited to non-transitory, tangible "storage" media, the term "readable medium" for a computer or machine refers to any medium that participates in providing instructions to a processor for execution.

[0267] Therefore, machine-readable media (such as computer-executable code) can take many forms, including but not limited to tangible storage media, carrier media, or physical transmission media. Non-volatile storage media include, for example, optical discs or disks, any storage device such as any one or more computers, such as those that can be used to implement the database shown in the accompanying drawings. Volatile storage media include dynamic memory, such as the main memory of such a computer platform. Tangible transmission media include coaxial cables; copper wires and optical fibers, including wires that form the bus within a computer system. Carrier transmission media can take the form of electrical or electromagnetic signals, or sound or light waves, such as those generated during radio frequency (RF) and infrared (IR) data communications. Therefore, common forms of computer-readable media include, for example: floppy disks, floppy hard disks, hard disks, magnetic tapes, any other magnetic media, CD-ROMs, DVDs or DVD-ROMs, any other optical media, punched card tapes, any other physical storage media with a perforated pattern, RAM, ROM, PROM and EPROM, FLASH-EPROM, any other memory chips or cassette tapes, carrier waves for transmitting data or instructions, cables or links for transmitting such carrier waves, or any other media from which a computer may read programming code and / or data. Many of these forms of computer-readable media may involve transmitting one or more sequences of one or more instructions to a processor for execution.

[0268] Computer system 1501 may include or communicate with an electronic display 1535, the electronic display including a user interface (UI) 1540 for providing, for example, images of biological components, barcodes, signals or measurement results of local parameters. Examples of UIs include, but are not limited to, graphical user interfaces (GUIs) and web-based user interfaces.

[0269] The methods and systems disclosed herein can be implemented using one or more algorithms. The algorithms can be implemented in software when executed by the central processing unit 1505. For example, the algorithms can identify biological components, detect barcodes, generate spatial modulation elements (e.g., masks), provide energy from an energy source, detect or measure local parameters using sensors, etc.

[0270] The following embodiments illustrate illustrative, non-limiting arrangements and combinations of the features disclosed herein. Other arrangements and combinations of features may also be considered.

[0271] Implementation Scheme 1: A method for processing chemical or biological materials, comprising: a) inputting (i) the chemical or biological material and (ii) one or more polymer precursors into a jetting device; b) supplying a stimulus to the jetting device to generate one or more polymer matrices, wherein the one or more polymer matrices comprise: i) a first gel structure from the one or more polymer precursors within the jetting device, wherein the first gel structure at least partially encapsulates the chemical or biological material, and wherein the first gel structure comprises a first internal region; and ii) a second gel structure adjacent to the first gel structure, wherein the second gel structure comprises a second internal region; and (c) degrading the first gel structure, the second gel structure, or a portion thereof, to generate a third internal region, wherein the third internal region comprises the first internal region of the first gel structure and the second internal region of the second gel structure. Implementation Scheme 2: The method of Implementation Scheme 1, wherein the first gel structure and the second gel structure each comprise one or more polymer matrix walls. Implementation Scheme 3: The method of Implementation Scheme 1, wherein the first gel structure and the second gel structure share a polymer matrix wall of one or more polymer matrix walls. Implementation Scheme 4: The method as described in Implementation Scheme 1, wherein the portion of the first gel structure, the second gel structure, or both degraded in (c) includes the shared polymer matrix wall.

[0272] Implementation Scheme 5: A method for processing chemical or biological materials, comprising: a) inputting (i) the chemical or biological material and (ii) one or more polymer precursors into a jetting device; and b) supplying a stimulus to the jetting device to generate one or more polymer matrices from the one or more polymer precursors within the jetting device, wherein the polymer matrix of the one or more polymer matrices at least partially encapsulates the chemical or biological material, and wherein the polymer matrix is ​​formed within less than 10 seconds after the one or more polymer precursors come into contact with the stimulus. Implementation Scheme 6: The method of Implementation Scheme 5, wherein the polymer matrix is ​​formed within less than 2 seconds after the one or more polymer precursors come into contact with the energy. Implementation Scheme 7: The method of Implementation Scheme 5, wherein the polymer matrix is ​​formed within less than 1 second after the one or more polymer precursors come into contact with the energy. Implementation Scheme 8: The method of Implementation Scheme 5, wherein the polymer matrix is ​​formed within less than 0.5 seconds or optionally within the range of about 2 seconds to about 4 seconds after the one or more polymer precursors come into contact with the energy.

[0273] Implementation Scheme 9: A method for treating chemical or biological materials, comprising: a) inputting (i) the chemical or biological material and (ii) one or more polymer precursors into a jetting device; b) supplying a stimulus to the jetting device to generate one or more polymer matrices from the one or more polymer precursors within the jetting device, wherein the polymer matrix of the one or more polymer matrices at least partially encapsulates the chemical or biological material; and c) performing one or more operations on the chemical or biological material within the polymer matrix, wherein the one or more operations include a time interval of at least one day, during which no more than 20% of the polymer matrix degrades. Implementation Scheme 10: The method of Implementation Scheme 9, wherein no more than 15% of the polymer matrix degrades during the time interval. Implementation Scheme 11: The method of Implementation Scheme 9, wherein no more than 10% of the polymer matrix degrades during the time interval. Implementation Scheme 12: The method of Implementation Scheme 9, wherein no more than 5% of the polymer matrix degrades during the time interval. Implementation Scheme 13: The method of Implementation Scheme 9, wherein no more than 1% of the polymer matrix degrades during the time interval. Implementation Scheme 14: The method as described in any one of Implementation Schemes 9-13, wherein the time interval includes at least three days. Implementation Scheme 15: The method as described in any one of Implementation Schemes 9-13, wherein the time interval includes at least five days. Implementation Scheme 16: The method as described in any one of Implementation Schemes 9-13, wherein the time interval includes at least seven days. Implementation Scheme 17: The method as described in any one of Implementation Schemes 9-13, wherein the time interval includes at least fourteen days.

[0274] Implementation Scheme 18: A method for treating chemical or biological materials, comprising: a) introducing (i) one or more chemical or biological materials and (ii) one or more polymeric precursors into a jetting device; b) supplying a stimulus to the jetting device to generate one or more polymeric matrices from the one or more polymeric precursors within the jetting device, wherein the one or more polymeric matrices at least partially encapsulate the one or more chemical or biological materials; and c) performing one or more operations on the one or more chemical or biological materials within the one or more polymeric matrices, wherein the one or more operations include a time interval of at least one day, wherein at least 80% of the one or more chemical or biological materials are viable during the time interval. Implementation Scheme 19: The method of Implementation Scheme 18, wherein at least 85% of the one or more chemical or biological materials are viable during the time interval. Implementation Scheme 20: The method of Implementation Scheme 18, wherein at least 90% of the one or more chemical or biological materials are viable during the time interval. Implementation Scheme 21: The method of Implementation Scheme 18, wherein at least 95% of the one or more chemical or biological materials are viable during the time interval. Implementation Scheme 22: The method of implementation scheme 18, wherein at least 99% of the one or more chemical or biological materials are viable during the time interval. Implementation Scheme 23: The method of any one of implementation schemes 18-22, wherein the time interval comprises at least three days. Implementation Scheme 24: The method of any one of implementation schemes 18-22, wherein the time interval comprises at least five days. Implementation Scheme 25: The method of any one of implementation schemes 18-22, wherein the time interval comprises at least seven days. Implementation Scheme 26: The method of any one of implementation schemes 18-22, wherein the time interval comprises at least fourteen days.

[0275] Embodiment 27: A method for processing chemical or biological materials, comprising: a) feeding (i) one or more chemical or biological materials and (ii) one or more polymer precursors into a jet device; and b) maintaining jet communication between the one or more chemical or biological materials and the polymer precursor for a time interval of at least ten minutes, wherein at least 80% of the one or more chemical or biological materials are viable during the time interval. Embodiment 28: The method of Embodiment 27, wherein at least 85% of the one or more chemical or biological materials are viable during the time interval. Embodiment 29: The method of Embodiment 27, wherein at least 90% of the one or more chemical or biological materials are viable during the time interval. Embodiment 30: The method of Embodiment 27, wherein at least 95% of the one or more chemical or biological materials are viable during the time interval. Embodiment 31: The method of Embodiment 27, wherein at least 99% of the one or more chemical or biological materials are viable during the time interval. Embodiment 32: The method of any one of Embodiments 27-31, wherein the time interval includes at least 30 minutes. Embodiment 33: The method of any one of embodiments 27-31, wherein the time interval includes at least one hour. Embodiment 34: The method of any one of embodiments 27-31, wherein the time interval includes at least two hours. Embodiment 35: The method of any one of embodiments 27-31, wherein the time interval includes at least three hours. Embodiment 36: The method of any one of embodiments 27-31, wherein the time interval includes at least one day. Embodiment 37: The method of any one of embodiments 27-36, further comprising supplying a stimulus to the jetting device to generate one or more polymer matrices from the one or more polymer precursors within the jetting device, wherein the one or more polymer matrices at least partially encapsulate the one or more chemical or biological materials. Embodiment 38: The method of any one of embodiments 1-26 and 37, wherein the stimulus comprises light. Embodiment 39: The method of any one of embodiments 1-26, 37, and 38, wherein the one or more polymer matrices comprise a polymeric form of cPEG monomers.

[0276] Implementation Scheme 40: The method of any one of Implementation Schemes 1-26 and 37-39, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (I): , Each instance of n is an integer from 0 to 500.

[0277] Implementation Scheme 41: The method of any one of Implementation Schemes 1-26 and 37-40, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (II): (II) where each instance of n is independently an integer from 0 to 500. Embodiment 42: The method as described in Embodiment 41, wherein n is between about 0 and about 100, or optionally n is between about 5 and about 50. Embodiment 43: The method as described in any one of Embodiments 1-42, wherein the one or more polymer precursors comprise at least 40 µL of solution.

[0278] Implementation Scheme 44: The method of any one of Implementation Schemes 1-26 and 37-43, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (III): (III), where n and m are independent integers from 0 to 500.

[0279] Implementation Scheme 45: The method of any one of Implementation Schemes 1-26 and 37-44, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (IV): , where each instance of n is an integer from 0 to 500 independently.

[0280] Implementation Scheme 46: The method of any one of Implementation Schemes 1-26 and 37-45, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising a structure (V): , where each instance of n is an integer from 0 to 500 independently.

[0281] Implementation Scheme 47: The method of any one of Implementation Schemes 1-26 and 37-46, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (VI): , where each instance of n is an integer from 0 to 500 independently.

[0282] Implementation Scheme 48: The method of any one of Implementation Schemes 1-26 and 37-47, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (VII): (VII)

[0283] Implementation Scheme 49: The method of any one of Implementation Schemes 1-26 and 37-48, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (VIII): , where n, m and p are independent integers from 0 to 500.

[0284] Implementation Scheme 50: The method of any one of Implementation Schemes 1-26 and 37-49, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising the structure (IX): (IX), where n is an integer from 0 to 500.

[0285] Implementation Scheme 51: The method of any one of Implementation Schemes 1-26 and 37-50, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (X): (X), where n and m are independent integers from 0 to 500.

[0286] Implementation Scheme 52: The method of any one of Implementation Schemes 1-26 and 37-51, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (XI): .

[0287] Implementation Scheme 53: The method of any one of Implementation Schemes 1-26 and 37-52, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (XII): , where each instance of n is an integer from 0 to 500 independently.

[0288] Implementation Scheme 54: The method of any one of Implementation Schemes 1-26 and 37-53, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising structure (XIII): , where each instance of n is an integer from 0 to 500 independently.

[0289] Embodiment 55: The method of any one of embodiments 1-26 and 37-54, wherein the one or more polymer matrices comprise a polymeric form of a monomer, the monomer comprising: an oligomer domain comprising three or more arms, wherein each arm of the oligomer domain comprises a degradable functional group and a crosslinkable functional group, wherein the crosslinkable functional group of one of the three or more arms is configured to crosslink with another crosslinkable functional group of another polymer precursor in response to a first stimulus to obtain the polymeric form of the monomer, and wherein the degradable functional group is configured to dissolve the polymeric form of the monomer in response to a second stimulus being cleaved. Embodiment 56: The method of embodiment 55, wherein the oligomer domain comprises four or more arms. Embodiment 57: The method of any one of embodiments 1-26 and 37-56, wherein the one or more polymer matrices comprise degradable functional groups. Embodiment 58: The method of Embodiment 58, wherein the degradable functional group comprises disulfide, β-thioether ester, amide hydroxymethyl and vicinal diol, vicinal diol, alginate backbone, dextran backbone, chitosan backbone, hyaluronic acid backbone, chondroitin sulfate backbone or carboxymethyl cellulose backbone or combinations thereof. Embodiment 59: The method of any one of Embodiments 1-26 and 37-58, wherein the one or more polymer matrices comprise a hydrogel. Embodiment 60: The method of Embodiment 60, wherein the hydrogel comprises a polymeric form of cPEG, cSEL-BTEEC, cSEL-DHEBA, cSEL-diol, cSEL-alginate, cSEL-dextran, cSEL-chitosan, cSEL-hyaluronic acid, cSEL-chondroitin sulfate or cSEL-cellulose or combinations thereof. Embodiment 61: The method of any one of Embodiments 1-26 and 37-60, further comprising contacting the one or more polymer matrices with a degradation agent configured to cleave the degradable functional groups. Embodiment 62: The method of Embodiment 62, wherein the degradation agent comprises DTT, TCEP, BME, GSH, DMEM, RPMI, PBS buffer, DMEM, RPMI, PBS buffer, (meta)periodate, alginate lyase (enzyme), dextranase, lysozyme and chitinase, hyaluronidase, chondroitinase or cellulase or combinations thereof. Embodiment 63: The method of any one of Embodiments 1-26 and 37-62, wherein the one or more polymer matrices comprise at least one β-thioether ester. Embodiment 64: The method of any one of Embodiments 1-26 and 37-63, wherein the one or more polymer matrices comprise PEG-m and one or more β-thioether esters. Embodiment 65: The method of Embodiment 63 or 64, wherein the β-thioether ester is formed by reacting an acrylate with a thiol.Embodiment 66: The method of any one of Embodiments 1-26 and 37-65, wherein the one or more polymer matrices are degraded overnight in the presence of a normally used cell culture medium, thereby providing a biocompatible degradation pathway for selective retention experiments. Embodiment 67: The method of any one of Embodiments 1-26 and 37-66, wherein the one or more polymer matrices comprise a Michael donor. Embodiment 68: The method of Embodiment 67, wherein the Michael donor is a PEG-thiol. Embodiment 69: The method of any one of Embodiments 1-26 and 37-68, wherein the one or more polymer matrices comprise a cSEL β-thioether ester having one β-thioether ester per arm. Embodiment 70: The method of any one of Embodiments 1-26 and 37-69, wherein the one or more polymer matrices comprise any material formed of PEG containing a Michael acceptor chain. Embodiment 71: The method of Embodiment 70, wherein the Michael acceptor chain comprises PEG-acrylamide, PEG-vinyl sulfone, PEG-maleimide, or PEG-carbonylacrylic acid, or any combination thereof. Embodiment 72: The method of any one of Embodiments 1-26 and 37-71, wherein the one or more polymer matrices are degraded by cleavage of disulfide bonds. Embodiment 73: The method of Embodiment 72, wherein the disulfide bonds are cleaved by one or more reducing agents. Embodiment 74: The method of Embodiment 73, wherein the one or more reducing agents comprise DTT, TCEP, BME, or GSH, or any combination thereof. Embodiment 75: The method of any one of Embodiments 1-26 and 37-74, wherein the one or more polymer matrices comprise one or more arms, each containing one or more amides. Embodiment 76: The method of any one of Embodiments 1-26 and 37-75, wherein the one or more polymer matrices are degraded by oxidative cleavage of vicinal diols by sodium periodate. Embodiment 77: The method of any one of Embodiments 1-26 and 37-76, further comprising: (a) using a caged workflow; (b) performing selective cell retention when used with cPEG or other hydrogels having an orthogonal degradation mechanism; or (c) performing long-term cell incubation using a reducing agent that does not affect cage stability in the cell culture medium. Embodiment 78: The method of any one of Embodiments 1-26 and 37-77, wherein the one or more polymer matrices comprise vicinal diol functionality that can be cleaved via oxidation of one or more hydroxyl groups. Embodiment 79: The method of any one of Embodiments 1-26 and 37-78, wherein the one or more polymer matrices are used to formulate a stable hydrogel.Embodiment 80: The method of any one of Embodiments 1-26 and 37-79, wherein the one or more polymer matrices comprise a polymeric form of a photocleavable 4-arm PEG-monomer. Embodiment 81: The method of any one of Embodiments 1-26 and 37-80, wherein the one or more polymer matrices are photodegraded via an o-nitrobenzyl moiety. Embodiment 82: The method of any one of Embodiments 1-26 and 37-81, wherein the one or more polymer matrices comprise a polymeric form of a coumarin-based photodegradable monomer. Embodiment 83: The method of any one of Embodiments 1-26 and 37-82, wherein the one or more polymer matrices comprise a 4-arm PEG-acrylamide containing one or more disulfides. Embodiment 84: The method of any one of Embodiments 1-26 and 37-83, wherein the one or more polymer matrices contain one or more cage disulfide bonds in the formation of hydrogel cages, wherein the hydrogel cages are degraded using light and a photoinitiator. Embodiment 85: The method of any one of Embodiments 1-26 and 37-84, wherein the one or more polymer matrices enable hydrogel formation, and wherein the hydrogel enables spatiotemporal control of hydrogel cage degradation, thereby enabling selective retention of cells with a single hydrogel formulation. Embodiment 86: The method of any one of Embodiments 1-26 and 37-85, wherein the one or more polymer matrices enable hydrogel formation, and wherein light and a photoinitiator as a reagent enable the hydrogel to selectively degrade cages and release their contents. Embodiment 87: The method of any one of the preceding embodiments, wherein the one or more polymer precursors enable the formation of a hydrogel up to 2 mm thick, and wherein the hydrogel can be degraded within 120 s with low light intensity. Embodiment 88: The method of any one of Embodiments 1-26 and 37-87, wherein upon exposure to light, photogenerated free radicals initiate multiple cleavage and disulfide exchange reactions, thereby allowing and promoting photodeformation, photowelding, and photodegradation of the one or more polymer matrices. Embodiment 89: The method of any one of Embodiments 1-26 and 37-88, wherein the one or more polymer precursors enable the formation of a hydrogel, wherein the hydrogel exhibits a chemical or physical change in response to an external stimulus. Embodiment 90: The method of any one of Embodiments 1-26 and 37-89, wherein the one or more polymer matrices comprise a photoinstantaneously unstable nitrobenzyl ester that cleaves upon photon absorption, thereby allowing the user to exogenously control the degradation of the one or more polymer matrices.Embodiment 91: The method of any one of Embodiments 1-26 and 37-90, wherein the one or more polymeric precursors enable hydrogel formation, and wherein the method further comprises controlling the network degradation of the hydrogel by means of the concentration of a photoinitiator supplied to the hydrogel. Embodiment 92: The method of any one of Embodiments 1-26 and 37-91, wherein the one or more polymeric precursors comprise thiol groups, which are quenched after the formation of the one or more polymeric matrices. Embodiment 93: The method of any one of Embodiments 1-26 and 37-92, wherein the polymeric matrix comprises a polymeric form of bisacrylamide. Embodiment 94: The method of any one of Embodiments 1-26 and 37-93, wherein the degradation of the one or more polymeric matrices depends on the type of encapsulated cells, the number of cells, or the fraction of disulfide moieties present in the one or more polymeric matrices. Embodiment 95: The method of any one of Embodiments 1-26 and 37-94, wherein the grafting efficiency of the one or more polymeric matrices is at least 90%. Embodiment 96: The method of any one of the preceding embodiments, wherein the one or more polymer precursors are in a solution with a pH of at least 6.0, 7.0, or 8.0. Embodiment 97: The method of any one of embodiments 1-26 and 37-96, wherein the one or more polymer precursors comprise a porogen, and wherein increasing the loading of the porogen results in an increased pore size of the one or more polymer matrices. Embodiment 98: The method of embodiment 97, wherein increasing the concentration of the porogen by 10% or 20% increases the diffusion rate by about 20%. Embodiment 99: The method of embodiment 98, wherein the increased pore size increases the diffusion rate of the fluorescently labeled antibody into the one or more polymer matrices. Embodiment 100: The method of embodiment 99, wherein the fluorescently labeled antibody is CD56 PE. Embodiment 101: The method of any one of the preceding embodiments, wherein the one or more polymer precursors are photopolymerized via photocrosslinking by UV light.

[0290] Implementation Scheme 102: A flow cell for sequencing genes, comprising: a plurality of cells, a plurality of nanopores and one or more polymer precursors; wherein the polymer matrix is ​​formed by the one or more polymer precursors within less than 10 seconds after the polymer precursors are contacted with energy.

[0291] Embodiment 103: A polymer precursor comprising: an oligomer domain having three or more arms, wherein each arm of the oligomer domain comprises a degradable functional group and a crosslinkable functional group, wherein the crosslinkable functional group of one of the three or more arms is configured to crosslink with another crosslinkable functional group of another polymer precursor in response to a first stimulus to obtain a polymeric form of the polymer precursor, and wherein the degradable functional group is configured to dissolve the polymeric form of the polymer precursor in response to a second stimulus being cleaved. Embodiment 104: A polymer precursor of embodiment 103, wherein the oligomer domain comprises four or more arms. Embodiment 105: A polymer precursor of embodiment 103 or 104, wherein the oligomer domain is hydrophilic. Implementation Scheme 106: A polymer precursor as described in any one of Implementation Schemes 103-105, wherein the oligomer domain comprises poly(ethylene glycol), poly(N-vinylpyrrolidone), poly(acrylic acid), poly(methacrylic acid), poly(vinyl alcohol), poly(L-lysine), poly(2-ethyl-2-oxazoline), poly(maleic acid), poly(vinyl phosphate), poly(acrylamide), poly(vinylamine), poly(ethylene oxide-co-propylene oxide), poly(N-isopropylacrylamide), poly(vinylphosphonic acid), poly(2-vinyl-1-methylpyridinium bromide), poly(N,N-diethylacrylamide), poly(N,N-dimethylacrylamide), poly(2-hydroxyethyl acrylate), poly(2-hydroxy methacrylate-co-methacrylic acid), poly(propylene oxide), poly(2-(dimethylamino)ethyl methacrylate), or poly(N-(2-hydroxypropyl)methacrylamide) or combinations thereof. Embodiment 107: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(ethylene glycol). Embodiment 108: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(N-vinylpyrrolidone). Embodiment 109: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(acrylic acid). Embodiment 110: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(methacrylic acid). Embodiment 111: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(vinyl alcohol). Embodiment 112: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(L-lysine). Embodiment 113: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(2-ethyl-2-oxazoline). Embodiment 114: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(maleic acid). Implementation Scheme 115: The polymer precursor as described in Implementation Scheme 106, wherein the oligomer domain comprises poly(vinyl phosphate).Embodiment 116: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(acrylamide). Embodiment 117: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(vinylamine). Embodiment 118: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(ethylene oxide-co-propylene oxide). Embodiment 119: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(N-isopropylacrylamide). Embodiment 120: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(vinylphosphonic acid). Embodiment 121: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(2-vinyl-1-methylpyridinium bromide). Embodiment 122: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(N,N-diethylacrylamide). Embodiment 123: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(N,N-dimethylacrylamide). Embodiment 124: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(2-hydroxyethyl acrylate). Embodiment 125: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(2-hydroxy methacrylate-co-methacrylic acid). Embodiment 126: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(propylene oxide). Embodiment 127: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(2-(dimethylamino)ethyl methacrylate). Embodiment 128: A polymer precursor as described in Embodiment 106, wherein the oligomer domain comprises poly(N-(2-hydroxypropyl)methacrylamide). Embodiment 129: A polymer precursor as described in any one of Embodiments 103-128, wherein the degradable functional group comprises disulfide, β-thioether ester, amide hydroxymethyl, vicinal diol, peptide, polysaccharide (e.g., alginate, dextran, chitosan, chondroitin), vicinal nitrobenzyl, coumarin, acetal, ketal, ester, orthoester, acid anhydride, imine, hydrazone, carbonate, phosphate, cinnamoyl, or benzoyl ether, or combinations thereof. Embodiment 130: A polymer precursor as described in Embodiment 129, wherein the degradable functional group comprises disulfide. Embodiment 131: A polymer precursor as described in Embodiment 129, wherein the degradable functional group comprises β-thioether ester. Embodiment 132: A polymer precursor as described in Embodiment 129, wherein the degradable functional group comprises amide hydroxymethyl. Embodiment 133: A polymer precursor as described in Embodiment 129, wherein the degradable functional group comprises vicinal diol. Implementation Scheme 134: The polymer precursor as described in Implementation Scheme 129, wherein the degradable functional group comprises a peptide.Embodiment 135: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises one or more polysaccharides. Embodiment 136: The polymer precursor of Embodiment 135, wherein the one or more polysaccharides comprises alginate. Embodiment 137: The polymer precursor of Embodiment 135, wherein the one or more polysaccharides comprises dextran. Embodiment 138: The polymer precursor of Embodiment 135, wherein the one or more polysaccharides comprises chitosan. Embodiment 139: The polymer precursor of Embodiment 135, wherein the one or more polysaccharides comprises chondroitin. Embodiment 140: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises o-nitrobenzyl. Embodiment 141: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises coumarin. Embodiment 142: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises acetal. Embodiment 143: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises ketal. Embodiment 144: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises an ester. Embodiment 145: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises an orthoester. Embodiment 146: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises an anhydride. Embodiment 147: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises animine. Embodiment 148: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises a hydrazone. Embodiment 149: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises a carbonate. Embodiment 150: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises a phosphate ester. Embodiment 151: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises a cinnamoyl group. Embodiment 152: The polymer precursor of Embodiment 129, wherein the degradable functional group comprises benzoin ether. Embodiment 153: The polymer precursor of any one of Embodiments 107-156, wherein the crosslinkable functional group comprises acrylate, methacrylate, acrylamide, methacrylamide, norbornel, styrene, vinyl ether, vinylpyrrolidone, vinyl ester, maleimide, allyl, alkyne, azide, thiol, olefin, or epoxide, or combinations thereof. Embodiment 154: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises acrylate. Embodiment 155: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises methacrylate.Embodiment 156: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises acrylamide. Embodiment 157: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises methacrylamide. Embodiment 158: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises norbornyl. Embodiment 159: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises styrene. Embodiment 160: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises vinyl ether. Embodiment 161: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises vinylpyrrolidone. Embodiment 162: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises vinyl ester. Embodiment 163: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises maleimide. Embodiment 164: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises an allyl group. Embodiment 165: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises an alkyne. Embodiment 166: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises an azide. Embodiment 167: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises a thiol. Embodiment 168: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises an olefin. Embodiment 169: The polymer precursor of Embodiment 157, wherein the crosslinkable functional group comprises an epoxide. Embodiment 170: The polymer precursor of any one of Embodiments 103-169, wherein the first stimulus comprises an energy source. Embodiment 171: The polymer precursor of Embodiment 170, wherein the energy source comprises a light source. Embodiment 172: The polymer precursor of Embodiment 171, wherein the wavelength of the light source is between 300-500 nm. Embodiment 173: A polymer precursor as described in any one of Embodiments 103-172, wherein the second stimulus comprises an energy source in the presence of a photoinitiator and in the absence of the polymer precursor. Embodiment 174: A polymer precursor as described in Embodiment 173, wherein the energy source comprises a light source. Embodiment 175: A polymer precursor as described in Embodiment 174, wherein the wavelength of the light source is between 300 and 500 nanometers (nm). Embodiment 176: A polymer precursor as described in Embodiment 175, wherein the wavelength of the light source is between 380 nm and 420 nm. Embodiment 177: A polymer precursor as described in any one of Embodiments 103-172, wherein the second stimulus comprises the introduction of a degradation agent.Embodiment 178: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises dithiothreitol (DTT). Embodiment 179: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises tris(2-carboxyethyl)phosphine (TCEP). Embodiment 180: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises 2-mercaptoethanol (BME). Embodiment 181: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises glutathione reductase (GSH). Embodiment 182: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises DMEM. Embodiment 183: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises RPMI. Embodiment 184: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises PBS buffer. Embodiment 185: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises sodium (meta)periodate. Embodiment 186: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises alginate lyase. Embodiment 187: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises dextranase. Embodiment 188: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises lysozyme. Embodiment 189: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises chitinase. Embodiment 190: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises hyaluronidase. Embodiment 191: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises chondroitinase. Embodiment 192: A polymer precursor as described in Embodiment 177, wherein the degrading agent comprises cellulase. Embodiment 193: A kit comprising: (a) a porogen; and (b) a polymer precursor described in any one of Embodiments 103-193. Embodiment 194: A kit as described in Embodiment 193, wherein the porogen comprises polyethylene glycol. Embodiment 195: A kit as described in Embodiment 193 or 194, wherein the ratio of the porogen to the polymer precursor by weight is between about 1:1 and 1:6. Embodiment 196: A kit as described in Embodiment 193 or 194, wherein the ratio of the porogen to the polymer precursor by weight is between about 1:2 and 1:5. Embodiment 197: A kit as described in Embodiment 193 or 194, wherein the ratio of the porogen to the polymer precursor by weight is between about 1:3 and 1:4. Embodiment 198: A kit as described in Embodiment 193 or 194, wherein the ratio of the porogen to the polymer precursor by weight is about 1:3.5.Embodiment 199: A kit as described in any one of Embodiments 193-198, wherein (a) the porogen and (b) the polymer precursor are contained in a single container. Embodiment 200: A kit as described in Embodiment 199, further comprising (c) a photoinitiator contained in another container. Embodiment 201: A kit as described in Embodiment 200, wherein the photoinitiator comprises one of lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), Irgacure 2959, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide (TPO) nanoparticles, 2,2'-azobis[2-methyl-N-(2-hydroxyethyl)promionamide] (VA-086), BAPO-Oli, BAPO-Ona, eosin Y, riboflavin, and combinations thereof.

[0292] Embodiment 202: The method of any one of Embodiments 1-4, wherein the chemical or biological material comprises a first cell in the first internal region and a second cell in the second internal region. Embodiment 203: The method of Embodiment 202, wherein the first cell comprises one of killer cells, activating cells, and inhibitory cells, and wherein the first cell interacts with the second cell after the degradation in (c). Embodiment 204: The method of any one of Embodiments 1-4, 202, and 203, wherein the chemical or biological material comprises polymer beads containing one or more oligonucleotides disposed thereon. Embodiment 205: The method of any of the preceding embodiments, wherein the chemical or biological material comprises one or more cells or components thereof. Embodiment 206: A polymer precursor of any one of Embodiments 103-192, wherein the oligomeric domain further comprises an amide moiety. Embodiment 207: A polymer precursor of any one of Embodiments 103-192 and 206, wherein the oligomeric domain further comprises a β-thioether moiety. Implementation Scheme 208: The method of any one of the preceding embodiments, wherein the one or more polymer precursors comprises: (a) a macromonomer, (b) a photoinitiator, and (c) a porogen. Implementation Scheme 209: The method of any one of embodiments 1-4, wherein the degradation described in (c) is caused by the cleavage of one or more disulfide bonds in the first gel structure, the second gel structure, or both.

[0293] Implementation Scheme 210: A method for treating chemical or biological materials, comprising: a) introducing (i) the chemical or biological material and (ii) one or more polymer precursors into a jetting device, wherein the one or more polymer precursors comprise a porogen, a macromonomer, and a photoinitiator; b) supplying an optical stimulus to the jetting device to generate two or more polymer matrices, wherein the two or more polymer matrices comprise: i) a first hydrogel structure formed within the jetting device, wherein the first hydrogel structure encapsulates the chemical or biological material, and wherein the first hydrogel structure includes a first internal region; and ii) a second hydrogel structure formed within the jetting device, wherein the second hydrogel structure encapsulates the chemical or biological material, and wherein the first hydrogel structure includes a second internal region; c) introducing a solution containing the photoinitiator into the jetting device; and d) supplying the optical stimulus to the jetting device to selectively degrade at least one of the first hydrogel structure or the second hydrogel structure.

[0294] Embodiment 211: The method of Embodiment 210, wherein feeding the solution containing the photoinitiator into the jet device includes removing the porogen and the macromonomer from the jet device. Embodiment 212: The method of Embodiment 210, wherein the macromonomer comprises a disulfide moiety, wherein two or more polymer matrices comprise the disulfide moiety, the disulfide moiety being configured to be cleaved upon exposure to the supplied optical stimulus. Embodiment 213: The method of any of the preceding embodiments, wherein the first hydrogel structure completely encapsulates the chemical or biological material. Embodiment 214: The method of any of the preceding embodiments, wherein the gel structure is a hydrogel structure. Embodiment 215: The method of any of the preceding embodiments, wherein the viable one or more biological materials comprise living cells. Embodiment 216: The kit of any one of Embodiments 193-198, wherein the polymer precursor is substantially free of a free thiol moiety, wherein the free thiol moiety is configured to degrade disulfide bonds. Implementation Scheme 217: A kit as described in Implementation Scheme 199, wherein the individual container is substantially free of the free thiol portion, wherein the free thiol portion is configured to degrade disulfide bonds. Implementation Scheme 218: A kit as described in Implementation Scheme 216 or 217, wherein the free thiol concentration is less than 5%, 1%, 0.1%, 0.01%, or 0.001%.

[0295] Example The following illustrative examples are representative embodiments of the apparatus and methods described herein and are not intended to be limiting in any way.

[0296] Example 1: Synthesis of cPEG macromonomer This example covers the synthesis of structure (I). The mixer was preheated to 35 °C using a 50 mL Smartblock tube. 957.4 mg of N,N'-bisacryloylcysteamine (BAC) was added to a 15 mL tube. 4000 µL of DMSO was added to the tube containing BAC. The tube was gently heated at 30 °C, vortexed, and sonicated to promote dissolution. 2400 mg of four-arm PEG-thiol (PEG-SH, CreativePEGworks, catalog number PSB-442, MW 5k) was added to a 50 mL Falcon tube. 22.60 mL of 1x PBS buffer (freshly prepared with nuclease-free water) was added to the PEG-SH in the Falcon tube, and then vortexed until dissolved. After the PEG-SH was dissolved, 17.8 µL of triethylamine was added to the PEG-SH solution. Using a syringe, rapidly add all the BAC / DMSO solution to a 50 mL tube containing PEG-SH and immediately vortex mix to prevent phase separation. Remove a 2 µL aliquot from the reaction mixture at time t=0 and monitor the reaction with an Ellman reagent test. Place the 50 mL tube in a mixer at 35 °C and shake at 850 rpm for 30 minutes. At time points t=15 min and 30 min, remove a 2 µL aliquot from the reaction mixture and monitor the reaction with an Ellman reagent test. After 30 min, reduce the temperature on the mixer to 25 °C and periodically remove the 2 µL aliquot, monitoring the reaction with an Ellman reagent test to determine completion (e.g., when the Ellman reading is within 5% of the PBS control). After completion, allow the reaction to continue for an additional 1 to 2 hours. Prepare a solution containing 266 mg of N-(2-hydroxyethyl)maleimide and 1 mL of 1x PBS buffer. Add the N-(2-hydroxyethyl)maleimide solution to the reaction mixture and stir for at least 30 minutes. Transfer the reaction mixture to 500-1000 Da Float-a-lyzer dialysis tubes and immerse them in 2-4 L beakers filled with MilliQ water. Perform four water changes according to the following schedule: 1) morning after the reaction (e.g., 9 am), 2) 3-4 hours after the first water change (e.g., 12-1 pm), 3) 2-4 hours after the second water change (e.g., 3-4 pm), and 4) 2-4 hours after the third water change (e.g., 5-7 pm). Transfer the dialysis solution to 50 mL centrifuge tubes (approximately 15-20 mL each) and freeze at -80 °C for at least 2 hours, then begin the freeze-drying process.

[0297] In some cases, photocleavable four-arm PEG macromonomers can be synthesized according to Scheme 1. In this case, the four-arm PEG macromonomer can be referred to as cPEG.

[0298] Option 1 In the cPEG reaction protocol above, (A) may include a 4-arm PEG thiol (5,000 g / mol), (B) may include N,N'-bis(acryloyl)cysteine, and (C) may include cPEG. The degradation mechanism may include the cleavage of disulfide bonds by a reducing agent such as DTT, TCEP, BME, GSH, etc. The product (e.g., cPEG) can be used in a caged workflow. In a further embodiment, when used with a second hydrogel that can be degraded via an orthogonal mechanism, the product (e.g., cPEG) may include selective cell retention.

[0299] Example 2: Procedure for monitoring free thiols cPEG (Ellman assay) A 4 mg / mL solution of Ellman's reagent (5,5′-dithiobis-(2-nitrobenzoic acid)) was prepared in a reaction buffer (e.g., 0.1 M sodium phosphate, pH 8.0, containing 1 mM EDTA). 125 µL of the reaction buffer was added to each well of a 96-well plate. Next, 2.5 µL of the 4 mg / mL Ellman's reagent solution was added to each well containing the reaction buffer. A 2 µL aliquot of the reaction mixture (e.g., at each time point) was removed and diluted with 142.4 µL of PBS to form a diluted aliquot of the reaction mixture. A 25 µL aliquot of the diluted reaction mixture was added to each well containing Ellman's reagent. After a 15-minute incubation period, the absorbance at 412 nm was measured in each well to assess the level of remaining thiol reactants in the reaction mixture.

[0300] Example 3: Synthesis of cSEL-diol macromonomer 616.1 µL of 1 M DTT solution, 11.4 µL of TEA, 1.258 mL of 1x PBS, and 208.2 µL of DMSO were combined in a 50 mL centrifuge tube and heated to 35 °C. A solution of 400.15 mg of four-arm PEG-acrylamide (CreativePEGworks, catalog number PSB-411, MW 5k) dissolved in 1874 µL of PBS was added dropwise over approximately 3 minutes to the DTT / TEA / PBS / DMSO mixture to form the reaction mixture. After 30 minutes, the reaction temperature was lowered to 25 °C. The addition of DTT to four-arm PEG-acrylamide was traced via an Ellman reaction (see Example 2). After 2 hours and 45 minutes, the reaction mixture was aspirated into a syringe and added dropwise over approximately 3 minutes at 35 °C to a solution of 175.48 mg of bisacrylamide in 727.5 µL of DMSO. The attachment of thiol-derived PEG-acrylamide to bisacrylamide was tracked using an Ellman assay until the thiol was completely consumed. Another Ellman assay was performed after 20 minutes to confirm the completion of the reaction. After 30 minutes, the temperature was lowered to 25 °C. 64.35 mg of N-(2-hydroxyhexyl)maleimide dissolved in 241.3 µL of PBS was added to the reaction mixture to quench any remaining thiol. The reaction mixture was then transferred to a 3.5 kDa molecular weight cutoff dialysis tube. After dialysis, the solution was freeze-dried to obtain cSEL-diol.

[0301] In some cases, photocleavable 4-arm PEG macromonomers can be synthesized as shown in Scheme 2. In this case, the 4-arm PEG macromonomer can be referred to as cSEL (β-thioether ester).

[0302] Option 2 Example 4: Synthesis of a 4-arm PEG-DTT-SH macromonomer A solution containing 150 mg of 4-arm PEG-acrylate (CreativePEGworks, catalog number PSB-423, MW5k) and 3.533 mL of PBS buffer was prepared. The 4-arm PEG-acrylate solution was added dropwise to a reaction vessel containing 0.23 mL of 1 M dithioerythritol in PBS buffer at 35 °C. After 30 minutes, the reaction temperature was lowered to 25 °C. The attachment of thiols from DTT to PEG-acrylate was monitored using an Ellman assay (see Example 2). After 4.5 hours, the reaction mixture was transferred to a 500–1000 Da MW cutoff dialysis tube for purification. After dialysis, 4-arm PEG-DTT-SH was obtained by lyophilization.

[0303] Example 5 - Synthesis of cSEL-BTEE macromonomer A solution containing 135.9 mg of a four-arm PEG-DTT-SH in 1.3 mL of PBS buffer was prepared and added dropwise to a PEG-diacrylate (Mn=250) solution containing 52 mg of PEG-diacrylate dissolved in 0.23 mL of DMSO over 3 minutes at 35 °C. Thiol conversion was tracked by Ellman assay (see Example 2). After 30 minutes, the reaction temperature was lowered to 25 °C, and the reaction was allowed to proceed for an additional 2 hours. The reaction was then quenched after 1 hour by adding 15.1 mg of N-(2-hydroxyethyl)maleimide dissolved in 56.8 µL of PBS, and the reaction was transferred to a 500–1000 Da molecular weight cutoff dialysis tube. After dialysis, the solution was lyophilized to obtain cSEL-BTEE.

[0304] In some cases, photocleavable 4-arm PEG macromonomers can be synthesized according to scheme 3. In this case, the 4-arm PEG macromonomer can be referred to as cSEL-BTEE.

[0305] Option 3 In some cases, photocleavable 4-arm PEG macromonomers can be synthesized as shown in Scheme 4. In this case, the 4-arm PEG macromonomer can be referred to as cSEL-BTEE.

[0306] Option 4 Example 6 - cSEL-DHEBA macromonomer An excess of N,N′-(1,2-dihydroxyethylene)bisacrylamide (DHEBA) solution (166.1577 mg DHEBA in 0.9 mL DMSO) was rapidly added to a solution of 4-arm PEG-thiol (CreativePEGworks, catalog number PSB-442, MW5k) (541.5 mg in 5.1 mL PBS buffer). The reaction mixture was allowed to be stirred at 35 °C for 30 min. The temperature was then lowered to 25 °C and allowed to proceed for 4 h. Thiol conversion was tracked via Ellman assay (see Example 2). After 4 h, the reaction was quenched by adding N-(2-hydroxyhexyl)maleimide (60 mg in 0.225 mL PBS). The quenching reaction was allowed to proceed for 30 min, and the solution was then transferred to a 500–1000 Da dialysis tube for purification. After dialysis, the resulting cSEL-DHEBA macromonomer solution was freeze-dried to obtain a solid material.

[0307] In some cases, photocleavable 4-arm PEG macromonomers can be synthesized according to scheme 5. In this case, the 4-arm PEG macromonomer can be referred to as cSEL-DHEBA.

[0308] Option 5 Example 7 - Photodegradable macromonomers based on coumarin In some embodiments, the photosynthetic gel is formed using a photoinitiator for free radical polymerization. In some embodiments, the photoinitiator includes Irgacure 2959, lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), or eosin Y (see, for example, Choi et al., Biotechniques, 66(1): 40-53 (2019)).

[0309] In some implementations, the hydrogel precursor includes 4-arm PEG, poly(ethylene glycol)-alkyne, such as cPEG (Example 1), cSEL-BTEE (Example 5), and cSEL-DHEBA (Example 6).

[0310] In some embodiments, the polymer precursor comprises PEG or multi-arm PEG. In some embodiments, the polymer precursor comprises an enzymatically degradable crosslinking agent. In some embodiments, such an enzymatically degradable crosslinking agent is degraded by esterases or peptidases. In some embodiments, the polymer precursor comprises a photodegradable crosslinking agent. In some embodiments, such a photodegradable crosslinking agent comprises a coumarin moiety.

[0311] In some embodiments, the photodegradable hydrogel is used in conjunction with the methods described herein, for example, because the photodegradation of the hydrogel chambers is selective and on-demand, allowing specific hydrogel chambers to degrade without affecting the unselected hydrogel chambers. In some embodiments, the hydrogel chambers undergo non-selective degradation, such that all hydrogel chambers in a given channel (or other container) degrade simultaneously. In some embodiments, such non-selective degradation is carried out using a cleaving agent that specifically cleaves unstable bonds in the hydrogel. For example, such cleaving agents include reducing agents. In some embodiments, such non-specific degradation is carried out using an enzyme that cleaves bonds or chemical elements in the hydrogel. For example, chemical elements include, but are not limited to, peptides, polysaccharides, and oligonucleotides.

[0312] In some cases, photocleavable 4-arm PEG macromonomers can be synthesized according to scheme 6. In this case, the 4-arm PEG macromonomer can be partially photodegraded via coumarin.

[0313] Option 6 Example 8 - Photodegradable monomers having an ortho-nitrobenzyl moiety In some embodiments, the photosynthetic gel is formed using a photoinitiator for free radical polymerization. In some embodiments, the photoinitiator includes Irgacure 2959, lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), or eosin Y (see, for example, Choi et al., Biotechniques, 66(1): 40-53 (2019)).

[0314] In some embodiments, the hydrogel precursor includes 4-arm PEG, poly(ethylene glycol)-alkyne (such as cPEG (Example 1), cSEL-BTEE (Example 5) and cSEL-DHEBA (Example 6)) and the photodegradable PEG macromonomer of Example 7.

[0315] In some embodiments, the polymer precursor comprises PEG or multi-arm PEG. In some embodiments, the polymer precursor comprises an enzymatically degradable crosslinking agent. In some embodiments, such an enzymatically degradable crosslinking agent is degraded by an esterase or peptidase. In some embodiments, the polymer precursor comprises a photodegradable crosslinking agent. In some embodiments, such a photodegradable crosslinking agent contains a nitrobenzyl group.

[0316] In some embodiments, the photodegradable hydrogel is used in conjunction with the methods described herein, for example, because the photodegradation of the hydrogel chambers is selective and on-demand, allowing specific hydrogel chambers to degrade without affecting unselected hydrogel chambers. In some embodiments, the hydrogel chambers undergo non-selective degradation, such that all hydrogel chambers in a given channel (or other container) degrade simultaneously. In some embodiments, such non-selective degradation is carried out using a cleaving agent that specifically cleaves unstable bonds in the hydrogel. For example, such cleaving agents include reducing agents. In some embodiments, such non-specific degradation is carried out using an enzyme that cleaves bonds or chemical elements in the hydrogel. For example, chemical elements include, but are not limited to, peptides, polysaccharides, and oligonucleotides.

[0317] In some cases, photocleavable 4-arm PEG macromonomers can be synthesized according to Scheme 7. In this case, the 4-arm PEG macromonomer can be photodegraded via the o-nitrobenzyl moiety.

[0318] Option 7 Example 9 - Cyclic lysis of hydrogel using sodium periodate This embodiment covers hydrogel chamber degradation using sodium periodate. Sodium periodate is capable of cleaving carbon-carbon bonds within vicinal diols, converting them to aldehydes via oxidative cleavage. A conventional schematic diagram of periodate-mediated vicinal diol cleavage is provided in Scheme 8 below.

[0319] Option 8 Hydrogel chambers were synthesized in three channels of a jet apparatus using a monomer of structure (XVII) containing four NaIO4-degradable vicinal diols. The chambers formed after irradiation for approximately 3–6 seconds in the presence of a photoinitiator. The chambers were washed and then stored in phosphate-buffered saline for approximately 15 minutes. The hydrogel chambers were then degraded at 25°C using the oxidant NaIO4. The hydrogel chambers degraded in less than two minutes.

[0320] Hydrogel chamber synthesis and NaIO4 degradation were then repeated in an additional jet apparatus at 4 °C. A stock solution of 50 mM NaIO4 in water was rapidly diluted to a concentration of 2.5 mM in cold PBS 1x and added to the channels of a caged jet apparatus. Chamber degradation was then measured using bright-field imaging while the jet apparatus was slowly warmed from 4 °C. Images of subsets of the measured chambers, taken at 5, 10, and 20 minutes after NaIO4 exposure, were displayed at [insert images here]. Figures 17A-17C As shown in the image, unlike the room-temperature NaIO4 degradation that occurs in less than two minutes, the hydrogel chamber remained slightly visible after 20 minutes.

[0321] Example 10 - Study on sodium periodate concentration in cage pyrolysis This embodiment addresses the degradation of a polymer matrix using a low sodium periodate concentration. Compared to Example 9, which presents a method for degrading a polymer matrix in less than 2 minutes, this embodiment covers a lower sodium periodate concentration that degrades the polymer matrix more slowly.

[0322] Hydrogel chambers containing four NaIO4-degradable vicinal diols were synthesized in three channels of a jet apparatus using a monomer of structure (XVII). The chambers were washed with 200 µL of phosphate-buffered saline. Then, after 15 minutes, 200 µL of NaIO4 in phosphate-buffered saline was added to each channel of the jet apparatus. The first channel received 1.25 mM NaIO4. The second channel received 0.613 mM NaIO4. The third channel received 0.306 mM NaIO4. Chamber degradation was then monitored using bright-field imaging.

[0323] Figure 18 This is a graph showing the degradation time (in minutes) of the hydrogel chambers as a function of NaIO4 concentration. The chamber exposed to 1.25 mM NaIO4 (channel 1) dissolved in approximately 2 minutes. The chamber exposed to 0.613 mM NaIO4 (channel 2) dissolved in approximately 6 minutes. The chamber exposed to 0.306 mM NaIO4 (channel 3) dissolved in approximately 20 minutes. The chambers in each channel were completely degraded.

[0324] Example 11 - Hydrogel Chamber Formation Rate This embodiment covers the photopolymerization rate of hydrogel chambers. The hydrogel chambers are formed using a polymer precursor comprising a structural (XVII) monomer and a type I photoinitiator, lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP). The polymer precursor is irradiated with blue light from a digital micromirror device (DMD) for one to three seconds to form the hydrogel chambers.

[0325] Then, bright-field images of the generated hydrogel chambers were collected. Figures 19A-19C These are top-view images of hydrogel chambers formed by irradiation with blue light for 1, 2, and 3 seconds, respectively. (Example) Figure 19A As shown, after 1 second of irradiation, faintly outlined hydrogel chambers were formed. However, as... Figures 19B-19C As shown, the hydrogel chambers formed after irradiation for 2 or 3 seconds exhibit higher densities. All three images show well-defined cylindrical morphologies in the hydrogel chambers, indicating that the polymer precursor can be rapidly photopolymerized in a spatially controlled manner.

[0326] Example 12 - Hydrogel chamber stability in the presence of adherent cells This embodiment covers the stability of hydrogel chambers. To test the ability of the hydrogel chambers to maintain their mechanical morphology, the hydrogel chambers were co-incubated with colonic fibroblasts; colonic fibroblasts are adherent cells that, after adhering to a surface, can exert mechanical stress by pulling on the hydrogel structure. The hydrogel chambers were formed in a jet apparatus by photopolymerization of a polymer precursor including a monomer comprising the four arms of structure (XVII). Each chamber was formed around a single colonic fibroblast. The jet apparatus was then washed, and the cells were incubated in culture medium for several days.

[0327] Bright-field images of the hydrogel chambers were collected immediately after hydrogel chamber synthesis. Figure 20A ) and collected 72 hours after hydrogel chamber synthesis ( Figure 20B After 72 hours, the cells increased in size and became translucent, indicating that they adhered to the bottom surface of the jet device. However, during this 72-hour period (from... Figures 20A to 20B The shape, width, and opacity of the chamber walls remained unchanged, indicating that colonic fibroblasts did not deform or degrade the hydrogel chambers.

[0328] Example 13 - Synthesis of macromonomers with disaccharide-cleavable functional groups This embodiment covers the synthesis of a multi-armed macromonomer containing a disaccharide-cleavable functional group in each arm. A summary of this synthesis is shown in Scheme 9. In the first step, a 4-armed PEG alkyne compound is reacted with an azide-modified disaccharide at room temperature in the presence of Cu(I) to link the disaccharide to the 4-armed PEG via click chemistry. The resulting compound is then reacted with acryloyl chloride to convert one or more disaccharide hydroxyl groups into a crosslinkable acrylate. Although Scheme 9 describes acryloyl chloride as undergoing nucleophilic substitution at the same hydroxyl group on each disaccharide unit, acryloyl chloride can be added to any hydroxyl group on each disaccharide. The number of disaccharide hydroxyl groups converted to acrylate can be controlled by modifying the ratio of acryloyl chloride to the disaccharide-modified 4-armed PEG. The resulting molecule is a crosslinkable macromonomer with a disaccharide-cleavable disaccharide in each of its four arms.

[0329] Option 9 Although preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. The invention is not intended to be limited to the specific embodiments provided herein. While the invention has been described with reference to the foregoing detailed description, the description and illustration of embodiments herein are not intended to be construed as limiting. Various variations, modifications, and substitutions will now occur to those skilled in the art without departing from the invention. Furthermore, it should be understood that all aspects of the invention are not limited to the specific depictions, configurations, or relative proportions described herein, which depend on various conditions and variables. It should be understood that various alternatives to the embodiments of the invention described herein can be used to implement the invention. Therefore, it is intended that the invention also cover any such alternatives, modifications, variations, or equivalents. The appended claims are intended to define the scope of the invention and are thereby intended to cover the methods and structures within the scope of these claims and their equivalents.

Claims

1. A polymer precursor comprising: An oligomer domain comprising three or more arms, wherein each arm of the oligomer domain comprises a degradable functional group and a crosslinkable functional group. The crosslinkable functional groups of the three or more arms are configured to crosslink with another crosslinkable functional group of another polymer precursor in response to a first stimulus, thereby obtaining a polymeric form of the polymer precursor, and the degradable functional groups are configured to dissolve the polymeric form of the polymer precursor in response to a second stimulus being cleaved.

2. The polymer precursor of claim 1, wherein the oligomer domain comprises four or more arms.

3. The polymer precursor of claim 1, wherein the polymer precursor comprises a compound of structure (I): .

4. The polymer precursor of claim 1, wherein the polymer precursor comprises a compound of structure (II): .

5. The polymer precursor of claim 1, wherein the polymer precursor comprises a structural (IV) compound: .

6. The polymer precursor of claim 1, wherein the polymer precursor comprises a structural (V) compound: .

7. The polymer precursor of claim 1, wherein the polymer precursor comprises a structural (VI) compound: .

8. The polymer precursor of claim 1, wherein the polymer precursor comprises a structure (XII) compound: .

9. The polymer precursor of claim 1, wherein the polymer precursor comprises a compound of structure (XIII): 。 10. The polymer precursor of claim 1, wherein the oligomeric domains are hydrophilic.

11. The polymer precursor of claim 10, wherein the oligomeric domain comprises poly(ethylene glycol), poly(N-vinylpyrrolidone), poly(acrylic acid), poly(methacrylic acid), poly(vinyl alcohol), poly(L-lysine), poly(2-ethyl-2-oxazoline), poly(maleic acid), poly(vinyl phosphate), poly(acrylamide), poly(vinylamine), poly(ethylene oxide-co-propylene oxide), poly(N-isopropylacrylamide), poly(vinylphosphonic acid), poly(2-vinyl-1-methylpyridinium bromide), poly(N,N-diethylacrylamide), poly(N,N-dimethylacrylamide), poly(2-hydroxyethyl acrylate), poly(2-hydroxy methacrylate-co-methacrylic acid), poly(propylene oxide), poly(2-(dimethylamino)ethyl methacrylate), or poly(N-(2-hydroxypropyl)methacrylamide) or combinations thereof.

12. The polymer precursor of claim 11, wherein the oligomer domain comprises poly(ethylene glycol).

13. The polymer precursor of claim 1, wherein the degradable functional group comprises disulfide, vicinal diol, β-thioether ester, amide hydroxymethyl, peptide, polysaccharide (e.g., alginate, dextran, chitosan, chondroitin), vicinal nitrobenzyl, coumarin, acetal, ketal, ester, orthoester, acid anhydride, imine, hydrazone, carbonate, phosphate, cinnamoyl or benzoyl ether, or combinations thereof.

14. The polymer precursor of claim 13, wherein the degradable functional group comprises a disulfide.

15. The polymer precursor of claim 13, wherein the degradable functional group comprises vicinal diol.

16. The polymer precursor of claim 1, wherein the crosslinkable functional group comprises acrylate, methacrylate, acrylamide, methacrylamide, norbornel, styrene, vinyl ether, vinylpyrrolidone, vinyl ester, maleimide, allyl, alkyne, azide, thiol, olefin or epoxide or a combination thereof.

17. The polymer precursor of claim 16, wherein the crosslinkable functional group comprises acrylate.

18. The polymer precursor of claim 17, wherein the crosslinkable functional group comprises acrylamide.

19. The polymer precursor of claim 1, wherein the first stimulus comprises an energy source.

20. The polymer precursor of claim 19, wherein the energy source comprises a light source.

21. The polymer precursor of claim 20, wherein the wavelength of the light source is between 300 and 500 nm.

22. The polymer precursor of claim 1, wherein the second stimulus comprises an energy source in the presence of a photoinitiator and in the absence of the polymer precursor.

23. The polymer precursor of claim 22, wherein the energy source comprises a light source.

24. The polymer precursor of claim 23, wherein the wavelength of the light source is between 300 and 500 nanometers (nm).

25. The polymer precursor of claim 24, wherein the wavelength of the light source is between 380 nm and 420 nm.

26. The polymer precursor of claim 1, wherein the second stimulus comprises the introduction of a degradation agent.

27. The polymer precursor of claim 26, wherein the degradation agent comprises a reducing agent.

28. The polymer precursor of claim 27, wherein the reducing agent comprises a thiol.

29. The polymer precursor of claim 28, wherein the thiol comprises glutathione, dithiothreitol (DTT), 2-mercaptoethanol (BME), or a combination thereof.

30. The polymer precursor of claim 26, wherein the degradation agent comprises an oxidant.

31. The polymer precursor of claim 30, wherein the oxidant comprises periodate.

32. The polymer precursor of claim 26, wherein the degradation agent comprises dithiothreitol (DTT), tris(2-carboxyethyl)phosphine (TCEP), 2-mercaptoethanol (BME), glutathione reductase (GSH), sodium (meta)periodate, alginate lyase, dextranase, lysozyme, chitinase, hyaluronidase, chondroitinase, cellulase, or combinations thereof.

33. A kit comprising: (a) a porogen; and (b) a polymer precursor as described in any one of claims 1-32.

34. The kit of claim 33, wherein the porogen comprises polyethylene glycol.

35. The kit of claim 33, wherein the ratio of the porogen to the polymer precursor by weight is between about 1:1 and 1:

6.

36. The kit of claim 33, wherein the ratio of the porogen to the polymer precursor by weight is between about 1:2 and 1:

5.

37. The kit of claim 33, wherein the ratio of the porogen to the polymer precursor by weight is between about 1:3 and 1:

4.

38. The kit of claim 33, wherein the ratio of the porogen to the polymer precursor by weight is about 1:3.

5.

39. The kit of claim 33, wherein (a) the porogen and (b) the polymer precursor are contained in a single container.

40. The kit of claim 33, further comprising (c) a photoinitiator contained in another container.

41. The kit of claim 40, wherein the photoinitiator comprises one of lithium phenyl-2,4,6-trimethylbenzoylphosphonate (LAP), Irgacure 2959, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide (TPO) nanoparticles, 2,2'-azobis[2-methyl-N-(2-hydroxyethyl)promionamide] (VA-086), BAPO-Oli, BAPO-Ona, eosin Y, riboflavin, and combinations thereof.

42. The polymer precursor of claim 1, wherein the oligomeric domain further comprises an amide moiety.

43. The polymer precursor of claim 1, wherein the oligomer domain further comprises a β-thioether moiety.

44. A method for treating chemical or biological materials, comprising: a) Inputting the chemical or biological material (i) and (ii) one or more polymeric precursors into the jet device, wherein the one or more polymeric precursors comprise a porogen, a macromonomer and a photoinitiator; b) Supplying optical stimulation to the jet device to generate two or more polymer matrices, wherein the two or more polymer matrices comprise: i) A first hydrogel structure formed within the jetting device, wherein the first hydrogel structure encapsulates a first chemical or first biological material, and wherein the first hydrogel structure includes a first internal region; and ii) A second hydrogel structure formed within the jetting device, wherein the second hydrogel structure encapsulates a second chemical or second biological material, and wherein the first hydrogel structure includes a second internal region; and c) Introduce the solution containing the photoinitiator into the jet device; d) Supply the optical stimulation to the jet device to selectively degrade at least one of the first hydrogel structure or the second hydrogel structure.

45. The method of claim 44, wherein feeding the solution containing the photoinitiator into the jetting apparatus includes removing the porogen and the macromolecular monomer from the jetting apparatus.

46. ​​The method of claim 44, wherein the macromonomer comprises a disulfide moiety, wherein two or more polymer matrices comprise the disulfide moiety, the disulfide moiety being configured to be cleaved upon exposure to the supplied optical stimulus.

47. The method of claim 44, wherein the first chemical or the first biological material is completely encapsulated within the first hydrogel structure.

48. The method of claim 44, wherein the first and second biological materials comprise living cells.

49. The kit of claim 33, wherein the polymer precursor is substantially free of free thiol moiety.

50. The kit of claim 33, wherein the individual container is substantially free of free thiol portion.

51. The kit of claim 50, wherein the concentration of the free thiol is less than 5%, 1%, 0.1%, 0.01%, or 0.001%.

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