# US 12,000,838 B2 · Reagents and methods for elemental mass spectrometry of biological samples > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,000,838 B2 - Document ID: US12000838B2 - Status: Granted - Kind: B2 - Application number: 17/050,716 - Filed: 2019-04-26 - Published: 2021-08-05 - Granted: 2024-06-04 - Earliest claimed: 2018-04-27 - Assignees: Standard BioTools Canada Inc. - Applicants: Fluidigm Canada Inc. - Inventors: Alexander V. Loboda; Olga Ornatsky; Vladimir Baranov; Nikita Zabinyakov; Anastasia Mavropoulos; Daniel Majonis; Bedilu Allo; Taunia Closson - Web page: https://patentgrep.com/patent/US12000838B2 ## CPC - G01N 33/6848: {Methods of protein analysis involving mass spectrometry} - H01J 49/0009: {Calibration of the apparatus} - C07K 1/13: Labelling of peptides - G01N 1/04: in the solid state, e.g. by cutting - G01N 1/28: Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00}) - G01N 1/4022: {by thermal techniques; Phase changes} - G01N 2001/045: {Laser ablation; Microwave vaporisation} - G01N 2458/15: Non-radioactive isotope labels, e.g. for detection by mass spectrometry - G01N 2560/00: Chemical aspects of mass spectrometric analysis of biological material - H01J 49/0409: {Sample holders or containers (containers for retaining a material to be analyzed, B01L3/50, for DNA, C12Q1/6834, for biological materials, G01N33/543)} - 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) - H01J: ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS (spark-gaps H01T; electric arc lamps H05B31/00; particle accelerators H05H) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract Embodiments of the present invention relate to reagents and their use for elemental imaging mass spectrometry of biological samples. The embodiments comprising methods for quantifying one or more analytes within a sample, comprising the steps of: (a) providing the sample, wherein the one or more analytes are immobilized to a sample carrier, wherein the sample has been labelled with one or more mass tags comprising one or more labelling atoms, (b) performing mass cytometry on the sample to determine the level of the one or more labelling atoms, wherein the level of the one or more labelling atoms corresponds to the copy number of the one or more analytes. ## Claims ### Claim 1 A sample carrier for mass cytometry, the sample carrier comprising: a planar surface, the planar surface including a first region for binding a sample; a polydopamine layer disposed on the planar surface; a self-assembled monolayer disposed on the polydopamine layer; a 3D polymer brush attached to the self-assembled monolayer; an element standard, the element standard comprising one or more elements or isotopes, wherein: the element standard comprises an element film applied to a second region of the planar surface using an adhesive, the element film is a polymer element film, and the first region does not overlap with the second region. ### Claim 2 (depends on 1) The sample carrier of claim 1, wherein the element standard comprises consistent abundance of the one or more elements or isotopes across the element standard. ### Claim 3 (depends on 1) The sample carrier of claim 1, wherein the element standard comprises separate regions, each with a different amount of the one or more elements or isotopes. ### Claim 4 (depends on 1) The sample carrier of claim 1, wherein different regions of the element standard may comprise a different combination of elements or isotopes. ### Claim 5 (depends on 1) The sample carrier of claim 1, wherein the element standard comprises reference particles of known elemental or isotopic composition, wherein the reference particles comprise metal elements or isotopes. ### Claim 6 (depends on 5) The sample carrier of claim 5, wherein the reference particles are optically resolvable. ### Claim 7 (depends on 5) The sample carrier of claim 5, wherein the reference particles comprise a metallic nanocrystal core and/or polymer surface. ### Claim 8 (depends on 1) The sample carrier of claim 1, wherein the element film is a polyester element film. ### Claim 9 (depends on 1) The sample carrier of claim 1, wherein the polymer element film comprises a long chain polymer that was mixed with a metal solution and volatile solvent prior to the solvent being evaporated. ### Claim 10 (depends on 1) The sample carrier of claim 1, wherein the element film comprises at least 10 different elemental isotopes, wherein the elemental isotopes have masses over 60 amu. ### Claim 11 (depends on 10) The sample carrier of claim 10, wherein the average number of the majority of elemental isotopes across the element film has a coefficient of variation (CV) of less than less than 10%. ### Claim 12 (depends on 1) A method of normalizing data obtained from a sample based on the element standard of the sample carrier of claim 1, wherein the sample comprises mass tags comprising labelling atoms, and wherein the method comprises analysing the sample by mass spectrometry. ### Claim 13 (depends on 12) The method of claim 12, further comprising using the element standard to generate a standard curve. ### Claim 14 (depends on 12) The method of claim 12, wherein the sample carrier is a sample slide comprising the sample, and further comprising applying the element film to the sample slide after tagging the sample with mass tags. ### Claim 15 (depends on 12) The method of claim 12, wherein normalization comprises quantification of the labelling atoms. ### Claim 16 (depends on 9) The sample carrier of claim 9, wherein the polymer is poly (methyl methacrylate) and the solvent is toluene. ### Claim 17 (depends on 9) The sample carrier of claim 9, further comprising the sample bound to the first region, wherein the sample is a biological sample. ### Claim 18 (depends on 9) The sample carrier of claim 9, wherein the element film is at least 100 square millimeters. ### Claim 19 (depends on 12) The method of claim 12, wherein the normalizing the data comprises analysing the element standard by mass spectrometry after analysing the sample. ### Claim 20 (depends on 1) A method comprising: binding the sample to the sample carrier of claim 1; ablating the element standard to form an ablation plume; transferring the ablation plume to a detector; and analysing the sample using the detector after transferring the ablation plume to the detector.