# US 2025/0076163 A1 · Method of sample preparation for maldi and automated system therefor > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2025/0076163 A1 - Document ID: US20250076163A1 - Status: Published application - Kind: A1 - Application number: 18/954,635 - Filed: 2024-11-21 - Published: 2025-03-06 - Earliest claimed: 2014-08-18 - Assignees: Becton, Dickinson and Company - Applicants: Becton, Dickinson and Company - Inventors: Timothy Wiles; John D. Mantlo - Web page: https://patentgrep.com/patent/US20250076163A1 ## CPC - G01N 1/38: Diluting, dispersing or mixing samples - G01N 1/2813: {Producing thin layers of samples on a substrate, e.g. smearing, spinning-on (G01N1/30 takes precedence)} - G01N 1/4022: {by thermal techniques; Phase changes} - G01N 1/44: Sample treatment involving radiation, e.g. heat - G01N 35/00722: {Communications; Identification} - G01N 35/1016: {Control of the volume dispensed or introduced} - G01N 15/075: by optical means - G01N 2001/4027: {evaporation leaving a concentrated sample} - H01J 49/0413: {for automated handling} - H01J 49/0418: {for laser desorption, e.g. matrix-assisted laser desorption/ionisation [MALDI] plates or surface enhanced laser desorption/ionisation [SELDI] plates} - H01J 49/0468: {with means for heating or cooling the sample} - H01J 49/40: Time-of-flight spectrometers (H01J49/36 takes precedence) - 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) ## Abstract Methods for preparing a biological sample for testing by Maldi where such methods are selected based on sample parameters. Maldi scores are obtained for a range of sample parameters (e.g. McFarland, dispense volume and number of dispenses). From the data, sample preparation parameters can be selected for a biological sample being prepared for Maldi testing. One sample preparation strategy uses multiple dispenses of sample with an intervening drying step, which yields more accurate Maldi scores, particularly for samples at the low range of McFarland values (e.g. below about 2). ## Claims ### Claim at position 1 1-36. (canceled) ### Claim 37 A non-transitory computer readable medium for causing a processor to control a method for preparing a biological sample for evaluation by Matrix-Assisted Laser Desorption Ionization Time of Flight mass spectrometry by: providing instructions to an automated apparatus, wherein the automated apparatus is adapted to obtain a biological sample from a culture media; and providing instructions to the automated apparatus to determine a volume of a diluent to be combined with the biological sample; and providing instructions to the automated apparatus to combine the volume of diluent with the biological sample to provide a diluted sample; providing instructions to the automated apparatus to measure a concentration of the diluted sample; wherein the processor receives the measured concentration of the diluted sample and determines if the measured concentration of the diluted sample is within a predetermined range, and, if the processor determines that the diluted sample is above a predetermined turbidity threshold in the predetermined range, providing instructions to the automated apparatus to apply a first predetermined volume of diluted sample to a substrate adapted to deliver a sample to a device configured to perform mass spectrometry; wherein, if the processor determines that a measured sample concentration of the diluted sample is below the predetermined turbidity threshold, providing instructions to the automated apparatus to: i) apply a second predetermined volume of the sample to a substrate adapted to deliver the sample to the device configured to perform mass spectrometry; ii) dry the sample; iii) apply a second application of the second predetermined volume of the sample over the dried sample on the substrate; and iv) dry the sample; v) apply a third application of the second predetermined volume of the sample of the dried sample on the substrate; vi) dry the sample; vii) apply a fourth application of the second predetermined volume of the dried sample on the substrate; viii) dry the sample; and providing instructions to the automated apparatus to apply a matrix over the applied sample, the matrix being adapted for use in the mass spectrometer. ### Claim 38 (depends on 37) The non-transitory computer readable medium of claim 37, wherein the processor provides instructions to cause a heater of the automated apparatus to dry the sample. ### Claim 39 (depends on 37) The non-transitory computer readable medium of claim 37, wherein the first predetermined volume and the second predetermined volume are the same. ### Claim 40 (depends on 37) The non-transitory computer readable medium of claim 37, wherein the second predetermined volume is about 3 μl. ### Claim 41 (depends on 37) The non-transitory computer readable medium of claim 37, wherein the first predetermined volume is about 3 μl. ### Claim 42 (depends on 37) The non-transitory computer readable medium of claim 37, wherein, if the processor determines that the diluted sample has a turbidity within the predetermined range, providing instructions to the automated apparatus to apply the diluted sample to the substrate in first and second applications with a drying step following each application. ### Claim 43 (depends on 37) The non-transitory computer readable medium of claim 37, wherein, if the processor determines that the diluted sample has a turbidity above the predetermined range, providing instructions to the automated apparatus to apply the diluted sample to the substrate in a first application with a drying step following the first application. ### Claim 44 (depends on 37) The non-transitory computer readable medium of claim 37, wherein the predetermined range of the measured sample concentration of the diluted sample is about 1 McFarland to about 2 McFarland. ### Claim 45 (depends on 37) The non-transitory computer readable medium of claim 37, wherein a range of the measured sample concentration of the diluted sample that is below the predetermined turbidity threshold is about 1 McFarland and below. ### Claim 46 (depends on 37) The non-transitory computer readable medium of claim 37, wherein the automated apparatus measures the concentration of the diluted sample with a nephelometer. ### Claim 47 A non-transitory computer readable medium for causing a processor to control a method for preparing a biological sample for evaluation by Matrix-Assisted Laser Desorption Ionization Time of Flight mass spectrometry by: providing instructions to an automated apparatus, wherein the automated apparatus is adapted to obtain a biological sample from a culture media; and providing instructions to the automated apparatus to determine a volume of a diluent to be combined with the biological sample; and providing instructions to the automated apparatus to combine the volume of diluent with the biological sample to provide a diluted sample; providing instructions to the automated apparatus to measure a concentration of the diluted sample; wherein the processor receives the measured concentration of the diluted sample and determines if the measured concentration of the diluted sample is within a predetermined range, and, if the processor determines that the diluted sample is above a predetermined turbidity threshold in the predetermined range, providing instructions to the automated apparatus to apply a first predetermined volume of diluted sample to a substrate adapted to deliver a sample to a device configured to perform mass spectrometry; wherein, if the processor determines that a measured sample concentration of the diluted sample is below the predetermined turbidity threshold, providing instructions to the automated apparatus to: i) apply a second predetermined volume of the sample to a substrate adapted to deliver the sample to the device configured to perform mass spectrometry; ii) dry the sample; iii) apply a second application of the second predetermined volume of the sample over the dried sample on the substrate; iv) dry the sample; and providing instructions to the automated apparatus to apply a matrix over the applied sample, the matrix being adapted for use in the mass spectrometer. ### Claim 48 (depends on 47) The non-transitory computer readable medium of claim 47, wherein the processor provides instructions to cause a heater of the automated apparatus to dry the sample. ### Claim 49 (depends on 47) The non-transitory computer readable medium of claim 47, wherein the first predetermined volume and the second predetermined volume are the same. ### Claim 50 (depends on 47) The non-transitory computer readable medium of claim 47, wherein the second predetermined volume is about 3 μl. ### Claim 51 (depends on 47) The non-transitory computer readable medium of claim 47, wherein the first predetermined volume is about 3 μl. ### Claim 52 (depends on 47) The non-transitory computer readable medium of claim 47, wherein, if the processor determines that the diluted sample has a turbidity within the predetermined range, providing instructions to the automated apparatus to apply the diluted sample to the substrate in first and second applications with a drying step following each application. ### Claim 53 (depends on 47) The non-transitory computer readable medium of claim 47, wherein, if the processor determines that the diluted sample has a turbidity above the predetermined range, providing instructions to the automated apparatus to apply the diluted sample to the substrate in a first application with a drying step following the first application. ### Claim 54 (depends on 47) The non-transitory computer readable medium of claim 47, wherein the predetermined range of the measured sample concentration of the diluted sample is about 1 McFarland to about 2 McFarland. ### Claim 55 (depends on 47) The non-transitory computer readable medium of claim 47, wherein a range of the measured sample concentration of the diluted sample that is below the predetermined turbidity threshold is about 1 McFarland and below. ### Claim 56 (depends on 47) The non-transitory computer readable medium of claim 47, wherein the automated apparatus measures the concentration of the diluted sample with a nephelometer.