Cell analysis method, cell analysis device, and cell analysis program

• US 9,405,958 B2
• Filed: 02/23/2012
• Issued: 08/02/2016
• Est. Priority Date: 04/28/2011
• Status: Active Grant

• Current Assignee: Hamamatsu Photonics KK
• Original Assignee: Hamamatsu Photonics KK
• Inventors: Kaneko, Kouichi, Sugiyama, Norikazu, Takeshima, Tomochika
• Primary Examiner(s): Shah, Utpal
• Assistant Examiner(s): Beg, Samah
• Application Number:
  US14/113,934
• Publication Number:
  US 20140064594A1
• Time in Patent Office: 1,622 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 33/5026 on cell morphology
• CPC Class Codes: G02B 21/14 affording illumination for phase-contrast observation
• CPC Class Codes: G06V 20/693 Acquisition
• CPC Class Codes: G06V 20/695 Preprocessing, e.g. image segmentation
• CPC Class Codes: G06V 20/698 Matching; Classification

Abstract

Provided is a cell analysis method in a cell analysis device D that uses an optical path length image of a cell colony formed of a large number of cells to analyze the cell colony, the method comprising: acquiring the optical path length image of the cell colony by an acquisition unit of the cell analysis device; extracting a circular shape corresponding to a cell nucleus of the cell in the acquired optical path length image by an extraction unit of the cell analysis device extracts; comparing an inner optical path length of the extracted circular shape and an outer optical path length of the extracted circular shape by a comparison unit of the cell analysis device extracts; and analyzing the cell colony based on the comparison result by analysis unit of the cell analysis device.

Claims

1. A cell analysis method in a cell analysis device that uses a quantitative optical path length image of a cell colony formed of a number of cells to analyze the cell colony, the method comprising:

acquiring the quantitative optical path length image of the cell colony by an acquisition unit of the cell analysis device;

extracting a circular shape corresponding to a cell nucleus of a cell in the acquired quantitative optical path length image by an extraction unit of the cell analysis device;

comparing an inner optical path length in an inner region of the extracted circular shape and an outer optical path length in an outer region of the extracted circular shape by a comparison unit of the cell analysis device; and

analyzing the cell colony based on the comparison result by an analysis unit of the cell analysis device to identify a cell with good quality, wherein the outer optical path length of the circular shape is larger than the inner optical path length of the circular shape for the cell with good quality.

2. The cell analysis method according to claim 1, wherein in the analyzing, when a number of cells per unit, which are determined as the cell with good quality, is equal to or greater than a threshold, the cell colony is determined as a cell colony with good quality.

3. The cell analysis method according to claim 2, further comprising:

specifying and displaying a region of the cell colony with good quality by display unit of the cell analysis device.

4. The cell analysis method according to claim 1, wherein the circular shape is a true circular shape.

5. The cell analysis method according to claim 4, wherein the true circular shape is a true circular shape having a predetermined diameter range.

6. The cell analysis method according to claim 1, wherein the circular shape is an elliptical shape.

7. The cell analysis method according to claim 1, wherein the cell is a stem cell.

8. A cell analysis system that uses a quantitative optical path length image of a cell colony formed of a number of cells to analyze the cell colony, the system comprising:

a microscope, comprising;

a stage that supports the cell colony; and

a light emission source that irradiates light onto the cell colony mounted on the stage;

an objective lens capable of viewing the irradiated cell colony on the stage;

a camera that captures at least one image of at least a portion of the irradiated cell colony; and

a cell analysis device, comprising;

memory configured to store the at least one image captured by the camera;

a processor communicatively coupled to the memory, the processor configured to;

acquire the quantitative optical path length image of the cell colony;

extract a circular shape corresponding to a cell nucleus of a cell in the acquired quantitative optical path length image;

compare an inner optical path length in an inner region of the extracted circular shape and an outer optical path length in an outer region of the extracted circular shape; and

analyze the cell colony based on the comparison result to identify a cell with good quality, wherein the outer optical path length of the circular shape is larger than the inner optical path length of the circular shape for the cell with good quality.

9. A non-transitory computer-readable medium containing program instructions for causing a computer to perform the method of:

in a cell analysis device that uses a quantitative optical path length image of a cell colony formed of a number of cells to analyze the cell colony, acquiring the quantitative optical path length image of the cell colony by an acquisition unit of the cell analysis device;

extracting a circular shape corresponding to a cell nucleus of a cell in the acquired quantitative optical path length image by an image extraction unit of the cell analysis device;

comparing an inner optical path length in an inner region of the extracted circular shape and an outer optical path length in an outer region of the extracted circular shape by a comparison unit of the cell analysis device; and

analyzing the cell colony based on the comparison result by an analysis unit of the cell analysis device to identify a cell with good quality, wherein the outer optical path length of the circular shape is larger than the inner optical path length of the circular shape for the cell with good quality.

10. A method for analyzing a cell based on quantitative optical path length image of the cell, the method comprising:

acquiring the optical path length image of the cell;

comparing a first quantitative optical path length of a first portion corresponding to a cell nucleus of the cell and a second quantitative optical path length of outside of the first portion;

and

determining that the cell is a cell with good quality based on the comparison result, wherein the second quantitative optical path length is larger than the first quantitative optical path length for the cell with good quality.

11. The method according to claim 10, further comprising:

extracting a shape corresponding to the cell nucleus in the acquired quantitative optical path length image, wherein the first portion comprises an inside of the shape.

12. The method according to claim 10, wherein the outside of the first portion corresponds to a cytoplasm of the cell.

13. The method according to claim 10, wherein the cell is a stem cell.

14. The method according to claim 13, wherein the stem cell is an embryonic stem (ES) cell or an induced pluripotent stem (iPS) cell.

15. The method according to claim 10, wherein the cell is included in a cell colony.

16. The method according to claim 10, wherein each of the first quantitative optical path length and the second quantitative optical path length is a phase difference or an optical thickness.

17. The method according to claim 10, wherein the quantitative optical path length image is captured by using a phase contrast microscope or a quantitative phase microscope.

18. An apparatus for analyzing a cell based on a quantitative optical path length image of the cell, the apparatus comprising:

a computer including a processor and configured to;

acquire the quantitative optical path length image of the cell,

compare a first optical path length of a first portion corresponding to a cell nucleus of the cell and a second optical path length of outside of the first portion, and

determine that the cell is a cell with good quality based on the comparison result, wherein the second optical path length is larger than the first optical path length for the cell with good quality.

19. The apparatus according to claim 18, wherein the computer extracts a shape corresponding to the cell nucleus in the acquired quantitative optical path length image, and wherein the first portion comprises an inside of the shape.

20. The apparatus according to claim 18, wherein the outside of the first portion corresponds to a cytoplasm of the cell.

21. The apparatus according to claim 18, wherein the cell is a stem cell.

22. The apparatus according to claim 21, wherein the stem cell is an embryonic stem (ES) cell or an induced pluripotent stem (iPS) cell.

23. The apparatus according to claim 18, wherein the cell is included in a cell colony.

24. The method according to claim 18, wherein each of the first quantitative optical path length and the second quantitative optical path length is a phase difference or an optical thickness.

25. The apparatus according to claim 18, wherein the quantitative optical path length image is captured by using a phase contrast microscope or a quantitative phase microscope.