Methods and devices for sample analysis

Patent metadata

Field	Value
Publication number	US-9562860-B1
Application number	US-201414309888-A
Country	US
Kind code	B1
Filing date	Jun 19, 2014
Priority date	Jun 19, 2013
Publication date	Feb 7, 2017
Grant date	Feb 7, 2017

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 cytometry apparatus for the analysis of stationary and flowing samples comprising:
a stage, wherein the stage is configured to receive and support a cytometry cuvette,
a cytometry cuvette, wherein the cytometry cuvette is supported by and is removable from the stage, wherein the cytometry cuvette comprises a sample plane, wherein the cytometry cuvette is configured to support therein a biological sample, and wherein a biological sample supported therein is supported on the sample plane of the cytometry cuvette;
an objective, wherein the objective comprises an entrance pupil and an exit pupil, and wherein the objective is optically coupled to the stage such that light from the biological sample traveling out of said sample plane may enter said entrance pupil and exit said exit pupil;
a sample coplanar light source disposed in substantially the same plane as the sample plane, wherein light from said sample coplanar light source is not in optical communication with said objective;
a sample coplanar optical sensor disposed substantially the same plane as the sample plane, wherein said sample coplanar optical sensor is not in optical communication with the objective;
a first detection train, wherein the first detection train comprises one or more image sensors in optical communication with said objective and configured for image analysis of stationary samples;
a second detection train, wherein the second detection train comprises one or more light detectors in optical communication with said objective and configured for the analysis of flowing samples;
an actuatable structure for directing light, said actuatable structure having a first position and a second position, wherein the actuatable structure is coupled to an actuation mechanism and, in said first position, the actuatable structure is configured to direct light from the exit pupil of the objective to the first detection train for the analysis of stationary samples and, in said second position, the actuatable structure is configured to direct light from the exit pupil of the second detection train for the analysis of flowing samples; and
an illumination train, wherein the illumination train comprises a light source in optical communication with said objective, 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 on a first location on the sample plane of the 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 cytometry cuvette.

2. A system for the analysis of stationary and flowing samples comprising:
a first cytometry cuvette, wherein the first cytometry cuvette is configured to receive and support a biological sample, wherein the first cytometry 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 configured to support a biological sample and upon which objects therein may settle;
a second cytometry cuvette, wherein the second cytometry cuvette is configured to receive and support a biological sample, wherein the second cytometry cuvette comprises a sample introduction port, a flow channel, and a sample collection area, wherein the sample introduction port, the flow channel, and the sample collection area are fluidically linked, wherein the flow channel comprises a bottom surface, wherein the bottom surface comprises a sample plane configured so that the biological sample and objects therein may flow over said bottom surface, and wherein the introduction port is configured to receive the biological sample wherein a biological sample received therein may flow the through flow channel, and collect in the sample collection area; and
a cytometry apparatus for the analysis of stationary and flowing samples, wherein the cytometry apparatus comprises:
a stage, wherein the stage is configured to receive and support the first cytometry cuvette and the second cytometry cuvette, and wherein the stage is configured to support one cytometry cuvette at a time;
an objective, wherein the objective comprises an entrance pupil and an exit pupil, and wherein the objective is optically coupled to the stage such that light from the biological sample traveling out of said sample plane may enter said entrance pupil and exit said exit pupil;
a sample coplanar light source disposed in substantially the same plane as the sample plane, wherein light from said sample coplanar light source is not in optical communication with said objective;
a sample coplanar optical sensor disposed substantially the same plane as the sample plane, wherein said sample coplanar optical sensor is not in optical communication with the objective;
a first detection train configured for image analysis of stationary samples, wherein the first detection train comprises one or more image sensors in optical communication with said objective;
a second detection train configured for the analysis of flowing samples, wherein the second detection train comprises one or more light detectors in optical communication with said objective;
an actuatable structure for directing light, said actuatable structure having, a first position and a second position, wherein the actuatable structure is coupled to an actuation mechanism and, in said first position, the actuatable structure is configured to direct light from the exit pupil of the objective to the first detection train configured for the analysis of stationary samples and, in said second position, the actuatable structure is configured to direct light from the exit pupil of the second detection train configured for the analysis of flowing samples; and
an illumination train, wherein the illumination train comprises a light source in optical communication with said objective, 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 on a first location on the sample plane of the second 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 second cytometry cuvette.

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

4. The apparatus of claim 1 , wherein the biological sample comprises multiple cells.

5. The apparatus of claim 1 , wherein light from the light source travels through the objective before reaching the sample plane of the cuvette.

6. The apparatus of claim 1 , wherein the cytometry cuvette is configured to support a stationary biological sample on the sample plane.

7. The apparatus of claim 1 , wherein the cytometry cuvette is configured to support a flowing biological sample on the sample plane.

8. The apparatus of claim 1 , wherein the cytometry cuvette comprises a sample introduction port, a flow channel, and a sample collection area, wherein the flow channel comprises the sample plane, wherein the sample introduction port, the flow channel, and the sample collection area are fluidically linked, and wherein the biological sample may be introduced in the introduction port, flow the through flow channel, and collect in the sample collection area.

9. The

Assignees

• Theranos Inc

Inventors

• Pangarkar Chinmay
• Mohan Karan

Classifications

• G01N15/1434

  Optical arrangements · CPC title

• G01N21/47

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

• G01N21/6428 Primary

  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

• G01N2201/06113

  Coherent sources; lasers · CPC title

• G01N21/6486 Primary

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