Methods and devices for sample analysis

Patent metadata

Field	Value
Publication number	US-10816475-B2
Application number	US-201916655750-A
Country	US
Kind code	B2
Filing date	Oct 17, 2019
Priority date	Jun 19, 2013
Publication date	Oct 27, 2020
Grant date	Oct 27, 2020

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

We claim:

1. 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 and wherein the cytometry apparatus comprises an illumination train, wherein the illumination train comprises a light source;
using an objective of the stage and a sample coplanar light source disposed to be substantially coplanar with the flow channel, wherein light from said sample light source is not in optical communication with the objective, wherein the objective is optically coupled to said stage such that light from the biological sample enters the objective;
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 output towards a first location in the flow channel and light of a second range of wavelengths from the light source is output towards a second location in the flow channel; and
detecting light emitted from a cell in the first location in the flow channel and light emitted from a cell in the second location in the flow channel.

2. 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 cytometry apparatus comprises an illumination train, wherein the illumination train comprises a light source;
using an objective of the stage and a sample coplanar light source disposed to be substantially coplanar with the flow channel, wherein light from said sample light source is not in optical communication with the objective, wherein the objective is optically coupled to said stage such that light from the biological sample enters the objective;
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 a 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 a bottom surface 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.

3. The method of claim 1 , wherein cells in the biological sample are white blood cells.

4. The method of claim 1 , wherein the first location and second location each have a diameter between 10 and 200 microns.

5. The method of claim 1 , 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.

6. The method of claim 1 , 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 an average diameter size of cells in the sample.

7. The method of claim 2 , further comprising a first detection train comprises at least 3 image sensors.

8. The method of claim 2 , further comprising a 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.

9. The method of claim 8 , 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.

10. The method of claim 1 , wherein the illumination train is further configured to focus light of a third range of wavelengths from the light source on a third location of the flow sample cuvette.

11. The method of claim 2 , 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.

12. The method of claim 2 , 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.

13. The method of claim 1 , 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.

14. 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;
using an objective of the stage and a sample coplanar light source disposed to be substantially coplanar with the flow channel, wherein light from said sample light source is not in optical communication with the objective, wherein the objective is optically coupled to said stage such that light from the biological sample enters the objective;
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 a 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 a cell in the first location in the flow channel and light emitted from a cell in the second location in the flow channel.

15. The method of claim 14 , wherein cells in the biological sample are white blood cells.

16. The method of claim 14 , wherein the first location and second location each have a diameter between 10 and 200 microns.

17. The method of claim 14 , wherein the first location and the

Assignees

• Theranos Ip Co Llc
• Labrador Diagnostics Llc

Inventors

• Pangarkar Chinmay
• Mohan Karan

Classifications

• G01N15/1459 Primary

  the analysis being performed on a sample stream · CPC title

• G01N21/47

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

• G01N2201/06113

  Coherent sources; lasers · 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

• G01N21/6486 Primary

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