IMAGE ANALYSIS AND MEASUREMENT OF BIOLOGICAL SAMPLES

• US 20140038206A1
• Filed: 07/25/2013
• Published: 02/06/2014
• Est. Priority Date: 07/25/2012
• Status: Active Grant

• Current Assignee: Labrador Diagnostics LLC (SoftBank Group Corp.)
• Original Assignee: Theranos Incorporated
• Inventors: Holmes, Elizabeth A., Pangarkar, Chinmay, Smith, Timothy, Young, Daniel L., Wasson, James R., Mohan, Karen, Anekal, Semartha
• Granted Patent: US 9,494,521 B2
• Time in Patent Office: Days
• US Class Current: 435/7.21
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 15/1012 Calibrating particle analysers; References therefor
• CPC Class Codes: G01N 2015/1014 Constitution of reference particles
• CPC Class Codes: G01N 2021/1738 Optionally different kinds of measurements; Method being valid for different kinds of measurement
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 21/05 Flow-through cuvettes G01N21/09 takes precedence; handling fluid samples G01N1/10
• CPC Class Codes: G01N 21/17 Systems in which incident light is modified in accordance with the properties of the material investigated where the material investigated is optically excited causing a change in wavelength of the incident light G01N21/63
• CPC Class Codes: G01N 21/27 using photo-electric detection G01N21/31 takes precedence; circuits for computing concentration logarithmic circuits G06G7/24; photometric circuits in general G01J
• CPC Class Codes: G01N 21/6428 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 Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 2201/061 Sources
• CPC Class Codes: G01N 2201/12 Circuits of general importance; Signal processing
• CPC Class Codes: G01N 2333/70589 CD45
• CPC Class Codes: G01N 2333/70596 Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
• CPC Class Codes: G01N 33/487 of liquid biological material
• CPC Class Codes: G01N 33/49 Blood chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80
• CPC Class Codes: G01N 33/5005 : involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/53 : Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/5308 : for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/56966 : Animal cells
• CPC Class Codes: G01N 33/56972 : White blood cells
• CPC Class Codes: G01N 33/582 : with fluorescent label
• CPC Class Codes: G02B 21/0076 : arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 21/088 : for both incident illumination and transillumination
• CPC Class Codes: G02B 21/125 : affording both dark- and bright-field illumination
• CPC Class Codes: G02B 21/16 : adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• CPC Class Codes: G02B 21/244 : using image analysis techniques
• CPC Class Codes: G02B 21/365 : Control or image processing arrangements for digital or video microscopes G02B21/361, G02B21/362 take precedence
• CPC Class Codes: G02B 7/09 : adapted for automatic focusing or varying magnification

Abstract

Methods, devices, systems, and apparatuses are provided for the image analysis of measurement of biological samples.

Claims

1. A system for analyzing a sample, the system comprising:

an illumination source;

a sample holder configured to receive light from the illumination source to provide both trans and epi illumination to the sample in the sample holder, wherein the illumination source is positioned to illuminate substantially only one side of the sample holder.

2. The system of any one of the foregoing claims, wherein total internal reflection is used to provide trans illumination.

3. The system of any one of the foregoing claims, wherein total internal reflection in the cuvette is used to provide trans illumination.

4. The system of any one of the foregoing claims, wherein the sample holder comprises at least one or more optically non-transmissive surfaces.

5. The system of any one of the foregoing claims, wherein the sample holder comprises a cuvette.

6. The system of any one of the foregoing claims, wherein the cuvette comprises two or more sample channels for holding sample.

7. The system of any one of the foregoing claims, wherein the cuvette has a circular horizontal, cross-sectional shape.

8. The system of any one of the foregoing claims, wherein the cuvette has a saw tooth vertical cross-sectional shape.

9. The system of any one of the foregoing claims, wherein the cuvette has a step-shaped vertical cross-sectional shape.

10. The system of any one of the foregoing claims, wherein the sample holder comprises an optically transmissive surface configured to positioned at a first location to be illuminated by the illumination source.

11. The system of any one of the foregoing claims further comprising an optical pathway from the illumination source to the sample holder wherein the pathway comprises a ringlight.

12. The system of any one of the foregoing claims further comprising an optical pathway from the illumination source to the sample holder wherein the pathway comprises an LED based ringlight.

13. The system of any one of the foregoing claims further comprising an optical pathway from the illumination source to the sample holder wherein the pathway comprises an laser based ringlight.

14. The system of any one of the foregoing claims further comprising compression device for holding the sample holder against in a desired location for illumination by the illumination source.

15. The system of any one of the foregoing claims further comprising an optically clear surface shaped to engage an optically clear surface of the sample holder.

16. The system of any one of the foregoing claims further comprising a detector configured to image only a portion of a channel in the sample holder.

17. The system of any one of the foregoing claims further comprising a detector configured to image an entire channel in the sample holder.

18. The system of any one of the foregoing claims wherein the sample holder is configured to hold the sample in a static, non-flowing manner during imaging.

19. The system of any one of the foregoing claims wherein during imaging, the sample holder is configured to hold one portion of the sample in a static, non-flowing manner and another portion in a flowing manner.

20. The system of any one of the foregoing claims wherein during imaging, the sample holder is configured to hold the sample in a flowing manner.

21. The system of any one of the foregoing claims wherein the sample remains separate from a detector in a fluid circuit fully confined in the sample holder, where the sample holder is movable relative to the detector.

22. A method for the measurement of a component of interest in cells of a cellular population in a sample, comprising:

a) obtaining a quantitative measurement of a marker present in cells of the cellular population in the sample;

b) based on the measurement of part a), determining, with the aid of a computer, an approximate amount of cells in the cellular population present in the sample;

c) based on the results of part b), adding an amount of a cell marker to add to the sample, wherein the marker binds specifically to the component of interest in cells of the cellular population and is configured to be readily detectable;

d) assaying cells in the sample for marker bound to the compound of interest; and

e) based on the amount of marker bound to the compound of interest, determining the amount of the component of interest in cells of the cellular population of the sample.

23. A method for focusing a microscope, comprising:

a) mixing a sample containing an object for microscopic analysis with a reference particle having a known size, to generate a mixture containing the sample and reference particle;

b) positioning the mixture of step a) into a light path of a microscope;

c) exposing the mixture of step a) to a light beam configured to visualize the reference particle; and

d) focusing the microscope based on the position of the reference particle within the mixture.

24. A method of identifying a cell in a sample containing a plurality of cells, comprising:

(a) assaying a cell of the plurality of cells for at least one of;

(i) the presence of a cell surface antigen;

(ii) the amount of a cell surface antigen;

or (iii) cell size;

(b) assaying the cell of (a) for at least one of;

(i) nuclear size;

or (ii) nuclear shape; and

(c) assaying the cell of (a) and (b) for quantitative cell light scatter,

wherein the combination of information from steps (a), (b), and (c) is used to identify the cell in the sample containing a plurality of cells