Methods and devices for sample analysis

Patent metadata

Field	Value
Publication number	US-2020116637-A1
Application number	US-201916655750-A
Country	US
Kind code	A1
Filing date	Oct 17, 2019
Priority date	Jun 19, 2013
Publication date	Apr 16, 2020
Grant date	—

Abstract

Methods and devices for cytometric analysis are provided. A cytometry apparatus is provided which may be used with a stationary sample cuvette for analysis of a stationary sample or with a flow sample cuvette for analysis of a flowing sample. The methods and devices provided herein may be used to perform cytometric analysis of samples under a wide range of experimental and environmental conditions.

First claim

1 - 17 . (canceled)

18 . A method of analyzing a biological sample, the method comprising:
moving a flow sample cuvette onto a stage of a cytometry apparatus, wherein the flow sample cuvette is configured to receive and support a biological sample, wherein the flow sample cuvette contains a flow channel through which a biological sample may flow, and wherein the flow channel comprises a bottom surface, wherein the bottom surface comprises a sample plane over which the biological sample and objects therein may flow, and wherein the cytometry apparatus comprises an illumination train, wherein the illumination train comprises a light source, wherein the illumination train is configured to provide at least a first illumination output and a second illumination output, and wherein for the first illumination output the illumination train is configured to focus light of a first range of wavelengths from the light source onto a first location on the sample plane of the flow cytometry cuvette and the illumination train is configured to focus light of a second range of wavelengths from the light source on a second location on the sample plane of the flow cytometry cuvette; loading a biological sample comprising multiple cells into the flow sample cuvette; flowing the biological sample through the flow channel of the flow sample cuvette, wherein during the flowing, light of a first range of wavelengths from the light source is focused on a first location in the flow channel and light of a second range of wavelengths from the light source is focused on a second location in the flow channel; and detecting light emitted from an cell in the first location in the flow channel and light emitted from an cell in the second location in the flow channel.

19 . A method of analyzing a biological sample, the method comprising:
dividing the biological sample into at least a first portion and a second portion, wherein both the first portion and the second portion contain multiple cells;
moving a flow sample cuvette onto the stage of a cytometry apparatus, wherein the flow sample cuvette is configured to receive and support a biological sample, wherein the flow sample cuvette contains a flow channel through which a biological sample may flow, and wherein the flow channel comprises a bottom surface, wherein the bottom surface comprises a sample plane over which the biological sample and objects therein may flow, and wherein the cytometry apparatus comprises an illumination train, wherein the illumination train comprises a light source, wherein the illumination train is configured to provide at least a first illumination output and a second illumination output, and wherein for the first illumination output the illumination train is configured to focus light of a first range of wavelengths from the light source onto a first location on the sample plane of the flow cytometry cuvette and the illumination train is configured to focus light of a second range of wavelengths from the light source on a second location on the sample plane of the flow cytometry cuvette;
loading the first portion of the biological sample into the flow sample cuvette;
flowing the first portion of the biological sample through the flow channel of the flow sample cuvette, wherein during the flowing, light of a first range of wavelengths from the light source is focused on a first location in the flow channel and light of a second range of wavelengths from the light source is focused on a second location in the flow channel; detecting light emitted from a cell in the first location in the flow channel and light emitted from an cell in the second location in the flow channel; removing the flow sample cuvette from the stage of the cytometry apparatus; moving a stationary sample cuvette to the stage of the cytometry apparatus, wherein the stationary sample cuvette is configured to receive and support a biological sample, wherein the stationary sample cuvette comprises a well for holding the biological sample in a stationary position, wherein the well comprises side walls and a bottom surface, and wherein the bottom surface of the well comprises a sample plane on which the biological sample and objects therein may settle; loading the second portion of the biological sample into the stationary sample cuvette; and
obtaining an image of some or all of the second portion of the biological sample in the stationary sample cuvette, wherein the image includes information from at least two cells in the second portion of the biological sample.

20 . The method of claim 18 , wherein the cells are white blood cells.

21 . The method of any of claim 18 , wherein the first location and second location each have a diameter between 10 and 200 microns.

22 . The method of any of claim 18 , wherein the first location and the second location each have a center, and wherein the center of the first location is separated from the center of the second location by a distance between 10 and 200 microns.

23 . The method of any of claim 18 , wherein the first location and the second location each have a center, and wherein the center of the first location is separated from the center of the second location by a distance of at least a average diameter size of cells in the sample.

24 . The method of any of claim 18 , wherein the first detection train comprises at least 3 image sensors.

25 . The method of any of claim 18 , wherein the second detection train comprises at least a first light detector, a second light detector, and a third light detector, wherein the detection train is configured such that the first light detector is configured to receive light of a first range of wavelengths, the second light detector is configured to receive light of a second range of wavelengths, and the third light detector is configured to receive light of a third range of wavelengths.

26 . The method of any of claim 18 , wherein the second detection train comprises a first emission filter before the first light detector, a second emission filter before second light detector, and a third emission filter before the third light detector.

27 . The method of any of claim 18 , wherein for first illumination output the illumination train is further configured to focus light of a third range of wavelengths from the light source on a third location on the sample plane of the cytometry cuvette.

28 . The method of any of claim 18 , wherein the first portion of the biological sample is loaded into the stationary sample cuvette before the stationary sample cuvette is moved onto the stage of the cytometry apparatus.

29 . The method of any of claim 18 , wherein the first portion of the biological sample is loaded into the stationary sample cuvette after the stationary sample cuvette is moved onto the stage of the cytometry apparatus.

30 . The method of any of claim 18 , wherein the first location in the flow channel and the second location in the flow channel are separated from each other by at least 10 and no more than 100 microns.

31 - 35 . (canceled)

36 . A method of analyzing a biological sample, the method comprising:
moving a flow sample cuvette onto the stage of a cytometry apparatus, wherein the flow sample cuvette is configured to receive and support a biological sample, wherein the flow sample cuvette contains a flow channel through which a biological sample may flow; loading a biological sample comprising multiple cells into the flow sample cuvette; flowing the biological sample through the flow channel of the flow sample cuvette, wherein during the flowing, light of a first range of wavelengths

Assignees

• Theranos Ip Co Llc

Inventors

• Pangarkar Chinmay
• Mohan Karan

Classifications

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• G01N21/6486 Primary

  Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence) · CPC title

• G01N21/47

  Scattering, i.e. diffuse reflection (G01N21/25, G01N21/41 take precedence {G01N21/55 takes precedence}) · CPC title

• G01N15/1459 Primary

  the analysis being performed on a sample stream · CPC title

• G01N21/4738

  Diffuse reflection (precedence is given to G01N21/55 - G01N21/57 if specular component is taken into consideration), e.g. also for testing fluids, fibrous materials · CPC title