# US 10,870,112 B2 · Selective delivery of material to cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,870,112 B2 - Document ID: US10870112B2 - Status: Granted - Kind: B2 - Application number: 16/145,865 - Filed: 2018-09-28 - Published: 2019-01-31 - Granted: 2020-12-22 - Earliest claimed: 2013-08-16 - Assignees: Massachusetts Institute of Technology - Applicants: Massachusetts Institute of Technology - Inventors: Armon R. Sharei; Viktor A. Adalsteinsson; Nahyun Cho; Robert S. Langer; J. Christopher Love; Klavs F. Jensen - Web page: https://patentgrep.com/patent/US10870112B2 ## CPC - B01L 3/502761: {specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules} - C12N 5/0634: {Cells from the blood or the immune system} - C12N 5/0693: {Tumour cells; Cancer cells} - C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation - C12Q 1/04: Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)} - G01N 15/1459: {the analysis being performed on a sample stream} - G01N 15/1484: {microstructural devices} - G01N 33/575: for cancer - B01L 2200/0652: Sorting or classification of particles or molecules - B01L 2300/08: Geometry, shape and general structure - B01L 2400/0487: fluid pressure, pneumatics - B82Y 30/00: Nanotechnology for materials or surface science, e.g. nanocomposites - C12M 23/16: {Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093)} - G01N 1/30: Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis} - G01N 15/01: specially adapted for biological cells, e.g. blood cells (investigating sedimentation of particle suspensions in blood G01N15/05) - G01N 2015/1006: {for cytology} - G01N 2015/1028: {Sorting particles} - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - 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) - B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} ## Abstract Isolating or identifying a cell based on a physical property of said cell can include providing a cell suspension; passing said suspension through a microfluidic channel that includes a constriction; passing the cell suspension through the constriction; and, contacting said cell suspension solution with a compound. The constriction can be sized to preferentially deform a relatively larger cell compared to a relatively smaller cell. ## Claims ### Claim 1 A method for delivering a compound into a cell based on a physical property of said cell comprising: providing a cell suspension comprising a first group of cells and a second group of cells, wherein the first group of cells have a relatively different size, diameter, and/or membrane stiffness than the second group of cells; passing said cell suspension through and out of a microfluidic channel that includes a constriction, said constriction being sized to deliver a compound to the first group of cells to a greater extent than the second group of cells, wherein passage through the constriction disrupts the cell membranes of the first group of cells such that the compound is delivered into the first group of cells to a greater extent than the second group of cells in the cell suspension that passes through the microfluidic channel; and contacting said cell suspension with the compound before the cell suspension is passed through the microfluidic channel that includes the constriction. ### Claim 2 (depends on 1) The method of claim 1, wherein the first group of cells have a relatively different size than the second group of cells. ### Claim 3 (depends on 1) The method of claim 1, wherein the first group of cells have a relatively different diameter than the second group of cells. ### Claim 4 (depends on 1) The method of claim 1, wherein the constriction has a diameter smaller than the size of cell in the cell suspension to which the compound is delivered. ### Claim 5 (depends on 1) The method of claim 1, wherein the constriction has a diameter smaller than the size of the smallest cell of the first group of cells or the smallest cell of the second group of cells. ### Claim 6 (depends on 1) The method of claim 1, wherein the constriction has a diameter smaller than the size of the largest cell of the first group of cells or the second group of cells. ### Claim 7 (depends on 1) The method of claim 1, wherein the cell suspension is further contacted with the compound after the suspension is passed through the microfluidic channel comprising the constriction. ### Claim 8 (depends on 1) The method of claim 1, wherein the cell suspension comprises whole blood. ### Claim 9 (depends on 1) The method of claim 1, wherein the cell suspension comprises peripheral blood mononuclear cells (PBMCs). ### Claim 10 (depends on 1) The method of claim 1, wherein the cell suspension comprises an erythrocycte-depleted population of peripheral blood cells. ### Claim 11 (depends on 1) The method of claim 1, wherein the compound has a molecular mass of 0.5 kDa to 5 MDa or a molecular mass of 3 kDa to 10 kDa. ### Claim 12 (depends on 1) The method of claim 1, wherein the compound comprises one or more of a protein, a nucleic acid, a detectable marker, an active biomolecule, and a toxin. ### Claim 13 (depends on 1) The method of claim 1, wherein the compound comprises a detectable marker. ### Claim 14 (depends on 1) The method of claim 1, wherein the constriction has a width from 4 μm-10 μm. ### Claim 15 (depends on 1) The method of claim 1, wherein the speed of the cells traversing the constriction ranges from 10 mm/s to 10 m/s. ### Claim 16 (depends on 1) The method of claim 1, further comprising applying a pressure to the cell suspension to drive the cell suspension through the constriction of the microfluidic channel. ### Claim 17 (depends on 1) The method of claim 1, wherein the first group of cells and/or the second group of cells comprise a cell selected from the group consisting of: tumor cells, PBMCs, and erythrocytes. ### Claim 18 (depends on 17) The method of claim 17, wherein the first group of cells are PBMCs. ### Claim 19 (depends on 17) The method of claim 17, wherein the PBMCs comprise leukocytes. ### Claim 20 (depends on 19) The method of claim 19, wherein the leukocytes comprise neutrophils, eosinophils, basophils, lymphocytes, and/or monocytes. ### Claim 21 (depends on 19) The method of claim 19, wherein the leukocytes are lymphocytes. ### Claim 22 (depends on 1) The method of claim 1, wherein the constriction has a length of 1 μm-100 μm. ### Claim 23 (depends on 1) The method of claim 1, wherein the constriction has 1-10 constrictions in series. ### Claim 24 (depends on 1) The method of claim 1, wherein the compound comprises a magnetic particle, a fluorescent particle, a fluorescent dye, a fluorescent protein, nucleic acid encoding a fluorescent protein, luciferase, or a compound that induces cell death. ### Claim 25 (depends on 1) The method of claim 1, wherein the compound comprises a detectable marker. ### Claim 26 (depends on 25) The method of claim 25, further comprising sorting the cells in the cell suspension based on uptake of the detectable marker, thereby isolating the first group of cells. ### Claim 27 (depends on 25) The method of claim 25, further comprising tagging the cells in the cell suspension with an antibody and sorting the cells in the cell suspension based on uptake of the detectable marker and/or antibody tagging, thereby isolating the first group of cells. ### Claim 28 (depends on 27) The method of claim 27, wherein the antibody binds a target on the first group of cells, and the cells are sorted by presence of the detectable marker and presence of the antibody tag, and wherein the antibody is a blood cell selective antibody. ### Claim 29 (depends on 27) The method of claim 27, wherein the antibody binds a target on the second group of cells, and cells are sorted by presence of the detectable marker and absence of the antibody tag, and wherein the antibody is a blood cell selective antibody. ### Claim 30 (depends on 1) The method of claim 1, wherein at least 10% more of the compound is delivered into the first group of cells compared to the second group of cells. ### Claim 31 (depends on 1) The method of claim 1, wherein the first group of cells have a relatively different membrane stiffness than the second group of cells. ### Claim 32 (depends on 1) The method of claim 1, wherein: the first group of cells and/or the second group of cells comprise a cell selected from the group consisting of: tumor cells, PBMCs, and erythrocytes; the first group of cells have a relatively different diameter than the second group of cells; the constriction has a diameter smaller than the diameter of a cell of the first group of cells in the cell suspension to which the compound is delivered; the constriction has a width from 4 μm-10 μm, a length of 1 μm-100 μm, and 1-10 constrictions in series; and the compound has a molecular mass of 0.5 kDa to 5 MDa.