# US 2019/0170625 A1 · Method of and apparatus for preparing samples for imaging or diffraction experiments under cryogenic conditions > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0170625 A1 - Document ID: US20190170625A1 - Status: Published application - Kind: A1 - Application number: 16/312,806 - Filed: 2017-06-22 - Published: 2019-06-06 - Earliest claimed: 2016-06-22 - Assignees: Universiteit Maastricht - Applicants: Universiteit Maastricht - Inventors: Franciscus Johannes Theresia Nijpels; Raimond Ravelli; Jacobus Peter Johannes Peters; Iglesias Carmen Lopez - Web page: https://patentgrep.com/patent/US20190170625A1 ## CPC - G01N 1/42: Low-temperature sample treatment, e.g. cryofixation - G01N 1/2813: {Producing thin layers of samples on a substrate, e.g. smearing, spinning-on (G01N1/30 takes precedence)} - H01J 37/20: Means for supporting or positioning the object or the material; Means for adjusting diaphragms or lenses associated with the support - F25D 2400/30: Quick freezing - H01J 2237/2001: Maintaining constant desired temperature - 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) - F25D: REFRIGERATORS; COLD ROOMS; ICE-BOXES; COOLING OR FREEZING APPARATUS NOT OTHERWISE PROVIDED FOR (refrigerated showcases A47F3/04; thermally-insulated vessels for domestic use A47J41/00; refrigerated vehicles, see the appropriate subclasses of classes B60 - B64; containers with thermal insulation in general B65D81/38; heat-transfer, heat-exchange or heat-storage materials, e.g. refrigerants, or materials for the production of heat or cold by chemical reactions other than by combustion C09K5/00; thermally-insulated vessels for liquefied or solidified gases F17C; air-conditioning or air-humidification F24F; refrigeration machines, plants, or systems F25B; cooling of instruments or comparable apparatus without refrigeration G12B) ## Abstract The invention relates to a method of and apparatus for preparing a sample for imaging or diffraction experiments under cryogenic conditions, comprising the steps of applying a sample to sample carrier, such as a film on a support, in particular a grid (3) comprising such a film on a support, or removing residual medium, typically liquid, from an incubated sample on a film on a support (3), and vitrifying the sample. The sample is vitrified by directing a jet (50, 51) of liquid coolant to the center of the film (3) and onto the sample. ## Claims ### Claim 1 A method of preparing a sample for imaging under cryogenic conditions or diffraction experiments under cryogenic conditions, comprising: applying a sample to a sample carrier, such as a film on a support, in particular a grid comprising such as a film on a support, or removing residual medium, typically liquid, from an incubated sample on a sample carrier, and vitrifying the sample, wherein the sample is vitrified by directing a jet of liquid coolant to the center of the sample carrier and onto the sample. ### Claim 2 (depends on 1) The method according to claim 1, wherein the time between completion of the application of the sample to the sample carrier and jet vitrification is less than a second. ### Claim 3 (depends on 1) The method according to claim 1, comprising simultaneously directing at least one jet of liquid coolant to each side of the support. ### Claim 4 (depends on 3) The method according to claim 3, wherein the jets are synchronized to within 0.5 ms from each other. ### Claim 5 (depends on 1) The method according to claim 1, wherein the velocity of the at least one jet of coolant is in a range from 1 to 50 m/s, preferably in a range from 1 to 20 m/s, and/or wherein the combined mass flow rate of the jets is in 0.1 to 100 L/min. ### Claim 6 (depends on 1) The method according to claim 1, wherein, after the sample has been vitrified, residues of the coolant are removed from the sample. ### Claim 7 (depends on 1) The method according to claim 1, wherein the sample, sample carrier and a gripper holding the sample carrier are advanced, after vitrification, into a coolant to cool the gripper and prevent heat from the gripper to undo vitrification. ### Claim 8 (depends on 1) The method according to claim 1, wherein, before applying the sample to the film, the film and the support are exposed to a plasma. ### Claim 9 (depends on 1) The method according to claim 1, wherein at least prior to application of the sample to the sample carrier the temperature of sample carrier is adjusted to a temperature sample carrier, at or above the dew point of the gas surrounding the sample carrier. ### Claim 10 (depends on 1) The method according to claim 1, wherein one or more of the above steps, preferably all steps are carried out in a closed chamber. ### Claim 11 An apparatus for preparing samples for imaging under cryogenic conditions or diffraction experiments under cryogenic conditions, comprising a holder for a sample carrier, such as a support carrying a film, an applicator for applying a sample to the sample carrier in the holder or a mechanism for removing residual medium from an incubated sample on the sample carrier, and wherein a nozzle for directing a jet of liquid coolant to the center of the sample carrier and onto the sample. ### Claim 12 (depends on 11) The apparatus according to claim 11, comprising at least one nozzle on each side of the support. ### Claim 13 (depends on 11) The apparatus according to claim 11, wherein the holder is movable between at least two positions, a first position at the applicator or the mechanism for removing residual medium and a further position in front of the nozzle(s). ### Claim 14 (depends on 11) The apparatus according to claim 11, comprising an inner vessel located beneath the nozzles and an outer vessel surrounding the inner vessel and wherein the nozzles are located at the discharge end of pipes that extend with their feed ends into the outer vessel. ### Claim 15 (depends on 11) The apparatus according to claim 11, comprising a cooling and/or heating element for the sample carrier, a temperature sensor and a humidity sensor, and a controller arranged to calculate the dew point of the gas surrounding the sample carrier and to adjust, by means of the element, the temperature of the sample carrier to a temperature around, at or above the dew point of the gas surrounding the sample carrier.