# US 9,434,937 B2 · Rapid cell purification systems > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,434,937 B2 - Document ID: US9434937B2 - Status: Granted - Kind: B2 - Application number: 14/004,145 - Filed: 2012-03-07 - Published: 2014-02-06 - Granted: 2016-09-06 - Earliest claimed: 2011-03-07 - Assignees: Accelerate Diagnostics, Inc. - Applicants: Steven W. Metzger; Kenneth Robert Hance - Inventors: Steven W. Metzger; Kenneth Robert Hance - Web page: https://patentgrep.com/patent/US9434937B2 ## CPC - C12N 13/00: Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves - B03C 5/005: {Dielectrophoresis, i.e. dielectric particles migrating towards the region of highest field strength} - C12M 47/12: {Purification (C12M47/04 takes precedence)} - C12Q 1/02: involving viable microorganisms - C12Q 1/24: Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms - G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)} - G01N 33/491: {by separating the blood components (G01N15/05 takes precedence)} - G02B 21/361: {Optical details, e.g. image relay to the camera or image sensor (G02B21/364 takes precedence; illumination details G02B21/06 and subgroups)} - B03C 2201/18: Magnetic separation whereby the particles are suspended in a liquid - B03C 2201/26: for use in medical or biological applications - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - C12N 7/02: Recovery or purification - C12N 2710/00051: Methods of production or purification of viral material - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - B03C: MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS (separating isotopes B01D59/00; combinations of magnetic or electrostatic separation with separation of solids by other means B03B, B07B) - 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} - 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) - G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS ## Abstract Methods and systems for purifying cells and/or viruses are provided. The sample is added to a well disposed in a medium. A potential is applied across the medium to cause the contaminants to enter one or more walls of the well, and retain the cells and/or viruses in the well. The cells and/or viruses can be removed from the well, and optionally adhered or fixed to a surface, or detected. In one embodiment, the cells and/or viruses may be retained in the well by embedding in the medium. The medium including the embedded cells and/or viruses may be excised or otherwise removed and transferred to a glass slide or other solid surface. ## Claims ### Claim 1 A method of purifying at least one of cells and viruses in a sample comprising: 1) adding a sample containing contaminants and at least one of cells and viruses to a well disposed in a porous medium; 2) applying an electrical potential across the medium to cause the contaminants to enter the medium through one or more walls of said well while retaining the at least one of the cells and viruses in said well; 3) reversing the electrical potential for a sufficient time to displace the at least one of the cells and viruses from the surface of the medium; and 4) removing the at least one of the cells and viruses from said well. ### Claim 2 (depends on 1) The method of claim 1, wherein said cells are microorganisms. ### Claim 3 (depends on 1) The method of claim 1, wherein said medium comprises at least one of a filter or a hydrogel. ### Claim 4 (depends on 3) The method of claim 3, wherein said hydrogel comprises at least one of polyacrylamide or agarose. ### Claim 5 (depends on 1) The method of claim 1, further comprising placing a buffer in contact with said medium. ### Claim 6 (depends on 5) The method of claim 5, wherein said buffer comprises histidine and tris(hydroxymethyl)aminomethane. ### Claim 7 (depends on 1) The method of claim 1, further comprising adding a chemical agent to the sample to increase a permeability of the medium to the contaminants. ### Claim 8 (depends on 1) The method of claim 1, further comprising applying a tangential flow to the medium to remove non-permeable contaminants from the surface of the medium. ### Claim 9 (depends on 1) The method of claim 1, wherein the electrical potential comprises an asymmetric alternating potential or a constant potential. ### Claim 10 (depends on 1) The method of claim 1, wherein removing the at least one of the cells and viruses from the well comprises removing at least a portion of the medium including a wall of the well having the at least one of the cells and viruses accumulated thereon. ### Claim 11 A method of purifying at least one of cells and viruses in a sample comprising: 1) adding a sample containing contaminants and at least one of cells and viruses to a well disposed in a porous medium; 2) applying an electrical potential across the medium to cause the contaminants to enter said medium through a surface of one or more walls of said well while retaining the at least one of the cells and viruses in said well; 3) providing a localization device to said medium, wherein the localization device draws the at least one of the cells and viruses in said well to a discrete location in the well thereby localizing the at least one of the cells and viruses; and 4) removing the at least one of the cells and viruses from said well; wherein the localization device is selected from at least one of a non-conductive material, a conductive material, and a discontinuous buffer system. ### Claim 12 (depends on 1) The method of claim 1, wherein applying the electrical potential and reversing the electrical potential are performed iteratively. ### Claim 13 A method of purifying at least one of cells and viruses in a sample comprising: adding a sample containing contaminants and at least one of cells and viruses to a well disposed in a medium; applying an electrical potential across the medium to cause the contaminants to enter said medium through a surface of one or more walls of said well while retaining the at least one of the cells and viruses in said well, wherein the electrical potential comprises a field polarity and the field polarity is switched according to a programmed sequence; and removing the at least one of the cells and viruses from said well. ### Claim 14 (depends on 1) The method of claim 1, further comprising mixing the sample. ### Claim 15 (depends on 14) The method of claim 14, wherein the sample is mixed by at least one of stirring the sample, repeatedly passing the sample through a pipette tip, and repeatedly forcing the sample through tubing connected to a syringe. ### Claim 16 (depends on 14) The method of claim 14, wherein the sample is mixed at least one of before application of the electrical potential, during application of the electrical potential, and after application of the electrical potential.