Cell analyzer, cell analyzer controlling method, and program

• US 10,604,731 B2
• Filed: 09/05/2017
• Issued: 03/31/2020
• Est. Priority Date: 03/06/2015
• Status: Active Grant

• Current Assignee: Sysmex Corporation
• Original Assignee: Sysmex Corporation
• Inventors: Masumoto, Kanako, Kubo, Takuya, Iwanaga, Shigeki, Okada, Masaya
• Primary Examiner(s): Wecker, Jennifer
• Application Number:
  US15/695,633
• Publication Number:
  US 20170362553A1
• Time in Patent Office: 938 Days
• CPC Class Codes: C12M 1/34 Measuring or testing with condition measuring or sensing means, e.g. colony counters
• CPC Class Codes: G01N 2021/6432 Quenching
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• 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 21/6456 Spatial resolved fluorescence measurements; Imaging
• 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: G06T 2207/30072 Microarray; Biochip, DNA array; Well plate
• CPC Class Codes: G06T 7/0012 Biomedical image inspection
• CPC Class Codes: G06V 20/693 Acquisition
• CPC Class Codes: G06V 20/698 Matching; Classification

Abstract

Provided is a cell analyzer including: a light source unit configured to apply light to test cells each containing first substances which are bound to first fluorescent dyes and which serve as an index for therapeutic strategy judgement; an image capturing unit configured to capture an image of fluorescence caused by the light; a processing unit configured to process the image obtained by the image capturing unit; and a display unit configured to display a process result obtained by the processing unit, wherein the processing unit obtains a first image by performing an inactivation process of quenching the first fluorescent dyes, an activation process of activating a part of the first fluorescent dyes that have been quenched, and an image capturing process of capturing, by means of the image capturing unit, an image of the fluorescence by applying light from the light source unit to each test cell; extracts bright points based on the first fluorescent dyes on the basis of the first image; classifies the extracted bright points into groups each corresponding to one first substance, thereby to obtain the number of the first substances in the test cell on the basis of the number of the classified groups; obtains therapy index information serving as an index for therapeutic strategy judgement, on the basis of the obtained number of the first substances; and causes the display unit to display the obtained therapy index information.

Claims

1. A cell analyzer comprising:

a light source unit configured to apply light to a test cell of which a first substance is stained with a first fluorescent dye;

an image capturing unit configured to capture an image of fluorescence caused by the light; and

a processing unit configured to process the image obtained by the image capturing unit,

wherein the processing unit is programmed to obtain a first image by performing an inactivation process of quenching the first fluorescent dye, an activation process of activating a part of the first fluorescent dye that have been quenched, and an image capturing process of capturing, by the image capturing unit, an image of the fluorescence by applying light from the light source unit to the test cells;

wherein said processing unit is programmed to extract bright points based on the first fluorescent dye on the basis of the first image; and

wherein said processing unit is programmed to classify the extracted bright points into groups each corresponding to one first substance, thereby to obtain the number of the first substances in the test cell on the basis of the number of the classified groups.

2. The cell analyzer of claim 1, wherein

the processing unit further obtains information regarding a distribution state of the first substances.

3. The cell analyzer of claim 1, wherein

the processing unit further obtains information regarding location of the first substances.

4. The cell analyzer of claim 1, wherein

each first substance is a gene that changes in a specific manner in a cell with a specific disease or a protein that changes in a specific manner in a cell with a specific disease.

5. The cell analyzer of claim 1, wherein

each first substance is a cancer marker gene or a cancer marker protein.

6. The cell analyzer of claim 1, wherein

the light source unit applies light to a test cell containing the first substance stained with the first fluorescent dye;

and second substance stained with a second fluorescent dye that are different from the first fluorescent dye, and

the processing unit further obtains a number of the second substance on the basis of a captured image of fluorescence generated from the second fluorescent dye.

7. The cell analyzer of claim 6, wherein

the processing unit further obtains information regarding location of the second substances.

8. The cell analyzer of claim 6, wherein

the processing unit obtains a therapy index information on the basis of a ratio between the number of the first substances and the number of the second substances.

9. The cell analyzer of claim 8, wherein

the processing unit determines presence/absence of change in the first substances on the basis of the ratio between the number of the first substances and the number of the second substances.

10. The cell analyzer of claim 6, wherein

the first substance is HER-2 gene contained in a nucleus of the test cell, and

the second substance is CEP17 contained in the nucleus of the test cell.

11. The cell analyzer of claim 6, wherein

the processing unit obtains a second image without performing an inactivation process of quenching the second fluorescent dye, and counts the number of the second substances in the test cell on the basis of the second image.

12. The cell analyzer of claim 6, wherein

the processing unit obtains a second image by performing an inactivation process of quenching the second fluorescent dye, an activation process of activating a part of the second fluorescent dye that have been quenched, and an image capturing process of capturing an image of the fluorescence by applying light from the light source unit to the test cell;

and counts the number of the second substances in the test cell on the basis of the second image.

13. The cell analyzer of claim 6, wherein

the processing unit obtains the first image and a second image by performing the inactivation process of quenching the first fluorescent dye and an inactivation process of quenching the second fluorescent dye, an activation process of activating a part of the first fluorescent dye that have been quenched and a part of the second fluorescent dye that have been quenched, and an image capturing process of capturing an image of the fluorescence by applying light from the light source unit to the test cell;

and counts the number of the second substances in the test cell on the basis of the second image.

14. The cell analyzer of claim 1, wherein

the processing unit

performs a first processing step in which, before the first image is obtained, a process of quenching and reactivating the first fluorescent dye is performed once, then the first image of the first fluorescent dyes is obtained, and a determination index indicating presence/absence of change in the first substances is obtained on the basis of the obtained first image;

when the change in the first substances obtained in the first processing step is negative, causes a display unit to display the determination index obtained in the first processing step, as therapy index information;

when the change in the first substances obtained in the first processing step is not negative, performs a second processing step in which

the first image is obtained by performing the inactivation process, the activation process, and the image capturing process, and

the number of the first substances is obtained by performing a process of extracting the bright points and classifying the bright points with respect to the obtained first image.

15. The cell analyzer of claim 1, wherein

the processing unit performs an activation process of activating the first fluorescent dye from a quenched state, by applying light having a predetermined wavelength.

16. The cell analyzer of claim 1, wherein

the processing unit further causes a display unit to display an image indicating a distribution state of the bright points of the first fluorescent dyes.

17. The cell analyzer of claim 1, wherein

the image capturing unit is configured to obtain, with respect to each first fluorescent dye, a captured image that allows identification of a position in an optical axis direction as well as a position on a two-dimensional plane at an image capture angle, and

the processing unit extracts bright points of a plurality of the first fluorescent dye on the basis of the positions on the two-dimensional plane and the positions in the optical axis direction, and classifies the extracted bright points into groups each corresponding to a first substance, thereby to obtain the number of the first substances.

18. The cell analyzer of claim 1, wherein

a nucleus of the test cell is stained by third fluorescent dyes, and

the processing unit identifies a region of the nucleus in the test cell on the basis of a captured image of fluorescence generated from the third fluorescent dye, and obtains the number of the first substances for each identified region.

19. A cell analyzer controlling method comprising:

obtaining a first image by performing

an inactivation process of quenching first fluorescent dye bound to one or more first substances in a test cell,

an activation process of activating a part of the first fluorescent dye that have been quenched, and

an image capturing process of capturing an image of fluorescence by applying light to the test cell;

extracting bright points based on the first fluorescent dye on the basis of the first image; and

classifying the extracted bright points into groups each corresponding to one first substance, thereby to obtain the number of the first substances in the test cell on the basis of the number of the classified groups.

20. A non-transitory computer-readable computer medium storing a program for causing a computer of a cell analyzer to perform operations, wherein said program is programmed to:

obtain a first image by performing an inactivation process of quenching a first fluorescent dye, an activation process of activating a part of the first fluorescent dye that have been quenched, and an image capturing process of capturing an image of the fluorescence;

extract bright points based on the first fluorescent dye on the basis of the first image; and

classify the extracted bright points into groups each corresponding to one first substance, thereby to obtain the number of the first substances in a test cell on the basis of the number of the classified groups.

21. A cell analyzer comprising:

a light source configured to irradiate light onto a test cell of which a first substance is stained with a first fluorescent dye;

an imaging device configured to capture an image of the test cell; and

a processing unit programmed to;

repeatedly quench and activate the first fluorescent dye in the test cell;

cause the imaging device to capture a plurality of images of the test cell under an irradiation by the light source while quenching and activating;

extract one or more bright points in the images; and

quantify a number of the first substance residing in the test cell based on the extracted bright points.

22. A cell analyzer comprising:

first and second light sources configured to irradiate light with different wavelengths onto a test cell of which a first substance is stained with a first fluorescent dye;

an imaging device configured to capture an image of the test cell; and

a processor programmed to;

cause the first light source to irradiate light onto the test cell to activate the first fluorescent dye in the test cell;

cause the second light source to irradiate light onto the test cell to quench the first fluorescent dye in the test cell;

cause the imaging device to capture a plurality of images of the test cell in the course of quenching during the irradiation by the second light source;

merge the images; and

quantify a number of the first substance residing in the test cell based on the merged image.