# US 10,481,068 B2 · Hydrogel particles with tunable optical properties > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,481,068 B2 - Document ID: US10481068B2 - Status: Granted - Kind: B2 - Application number: 15/895,307 - Filed: 2018-02-13 - Published: 2018-09-06 - Granted: 2019-11-19 - Earliest claimed: 2012-04-06 - Assignees: Slingshot Biosciences, Inc. - Applicants: Slingshot Biosciences - Inventors: Jeffrey Kim; Oliver Liu; Jeremy Agresti; Anh Tuan Nguyen - Web page: https://patentgrep.com/patent/US10481068B2 ## CPC - G01N 15/1012: {Calibrating particle analysers; References therefor} - G01N 15/14: Optical investigation techniques, e.g. flow cytometry - G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)} - G01N 15/149: specially adapted for sorting particles, e.g. by their size or optical properties - G01N 2015/1006: {for cytology} - G01N 2015/1014: {Constitution of reference particles} - G01N 2015/1415: {Control of particle position} - G01N 2021/6439: {with indicators, stains, dyes, tags, labels, marks} - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The present disclosure relates to compositions comprising a hydrogel particle with optical properties substantially similar to the optical properties of a target cell, and methods for their use. ## Claims ### Claim 1 A method, comprising: calibrating a cytometric device for analysis of a human target cell, the calibrating including: inserting hydrogel particles having optical scatter properties at least 70% similar to optical scatter properties of the human target cell into the cytometric device, a physical property of the hydrogel particles being closer in value to a corresponding physical property of the human target cell than a corresponding physical property of polystyrene beads having substantially the same average diameter as the hydrogel particles, the optical scatter properties of the hydrogel particles provided by: a co-monomer, a chemical side-group, an encapsulated material, a colloidal silica, or a ratio of acrylamide to bi-acrylamide, measuring the physical property of the hydrogel particles using the cytometric device, and calibrating the cytometric device based on the measurements of the hydrogel particles; and analyzing the human target cell with the calibrated cytometric device. ### Claim 2 (depends on 1) The method of claim 1, wherein the hydrogel particles are analyzed simultaneously with the human target cell. ### Claim 3 (depends on 1) The method of claim 1, wherein the hydrogel particles are measured by the cytometric device as an external standard. ### Claim 4 (depends on 1) The method of claim 1, wherein the hydrogel particles are configured to be manipulated by an end-user to act as a process control. ### Claim 5 (depends on 1) The method of claim 1, wherein the hydrogel particles contain measurable nucleotide sequences, the calibrating further including: measuring the nucleotide sequences of the hydrogel particles. ### Claim 6 (depends on 1) The method of claim 1, further comprising sorting cells of the analyzed human target cell using the cytometric device. ### Claim 7 (depends on 1) The method of claim 1, wherein the hydrogel particles have an average refractive index of greater than 1.7. ### Claim 8 (depends on 1) The method of claim 1, wherein the hydrogel particles have an average diameter of less than 70 mm. ### Claim 9 (depends on 1) The method of claim 1, wherein no more than 10% of the hydrogel particles have a diameter exceeding the average diameter of the hydrogel particles by more than 10%. ### Claim 10 (depends on 1) The method of claim 1, wherein the hydrogel particles do not include agarose. ### Claim 11 (depends on 1) The method of claim 1, wherein the optical scatter properties of the hydrogel particles are defined by: a co-monomer of the hydrogel particles; or the hydrogel particles being functionalized with a stoichiometrically-tuned ratio of at least one chemical side-group and at least one secondary label. ### Claim 12 (depends on 1) The method of claim 1, wherein the hydrogel particles include a polymerized monomer, the monomer including: lactic acid, glycolic acid, acrylic acid, 1-hydroxyethyl methacrylate, ethyl methacrylate, propylene glycol methacrylate, acrylamide, N-vinylpyrrolidone, methyl methacrylate, glycidyl methacrylate, glycol methacrylate, ethylene glycol, or fumaric acid. ### Claim 13 (depends on 1) The method of claim 1, wherein the hydrogel particles include a monofunctional monomer, the monofunctional monomer including: acrylamide, methacrylamide, an N-alkylacrylamide, an N-alkylmethacrylamide, an N,N-dialkylacrylamide, an N-[(dialkylamino)alkyl]acrylamide, an N-[(dialkylamino)alkyl]methacrylamide, a (dialkylamino)alkyl acrylate, or a (dialkylamino)alkyl methacrylate. ### Claim 14 (depends on 1) The method of claim 1, wherein the hydrogel particles include a bifunctional monomer, the bifunctional monomer including: allyl amine, allyl alcohol, allyl isothiocyanate, allyl chloride, or allyl maleimide. ### Claim 15 (depends on 1) The method of claim 1, wherein the hydrogel particles include a bifunctional acrylic monomer. ### Claim 16 A method of calibrating a measurement device for analysis of a human target cell, the method comprising: a) inserting into the measurement device hydrogel particles having optical scatter and/or biochemical properties at least 70% similar to optical scatter and/or biochemical properties of a human target cell type, a material modulus property of the hydrogel particles being closer in value to a corresponding material modulus property of the human target cell than a corresponding material modulus property of polystyrene beads having substantially the same average diameter as the hydrogel particles, the optical scatter properties of the hydrogel particles provided by: a co-monomer, a chemical side-group, an encapsulated material, a colloidal silica, or a ratio of acrylamide to bi-acrylamide; b) measuring the properties of the hydrogel particles using the measurement device; and c) calibrating the measurement device based on the measurements of the hydrogel particles. ### Claim 17 (depends on 16) The method of claim 16, wherein the measurement device is not adjusted between measuring the optical scatter properties of the hydrogel particles and a subsequent human target cell analysis. ### Claim 18 (depends on 16) The method of claim 16, wherein the hydrogel particles are substantially optically and/or biochemically indistinguishable from cellular matter as measured by the measurement device. ### Claim 19 (depends on 16) The method of claim 16, wherein the hydrogel particles are used as an internal standard and are analyzed simultaneously with cellular matter. ### Claim 20 (depends on 16) The method of claim 16, wherein the hydrogel particles contain nucleotide sequences, the method further including measuring the nucleotide sequences of the hydrogel particles. ### Claim 21 (depends on 16) The method of claim 16, further comprising: analyzing a human target cell with the calibrated measurement device. ### Claim 22 (depends on 21) The method of claim 21, further comprising: separating cells of the analyzed human target cell using the measurement device. ### Claim 23 (depends on 21) The method of claim 21, further comprising determining mass spectrometry properties of the human target cell. ### Claim 24 (depends on 16) The method of claim 16, wherein the hydrogel particles contain nanoparticles. ### Claim 25 (depends on 16) The method of claim 16, wherein the hydrogel particles contain nanoparticles of PMMA. ### Claim 26 (depends on 16) The method of claim 16, wherein the hydrogel particles contain quantum dots. ### Claim 27 (depends on 16) The method of claim 16, wherein the hydrogel particles contain isotope-labeled proteins, antibodies, or a combination thereof.