Microscope cell staining observation system, method, and computer program product

• US 9,110,305 B2
• Filed: 11/19/2010
• Issued: 08/18/2015
• Est. Priority Date: 02/26/2010
• Status: Active Grant

• Current Assignee: Evident Corporation, Japanese Foundation For Cancer Research
• Original Assignee: Japanese Foundation For Cancer Research, Olympus Corporation
• Inventors: Yamada, Tatsuki, Ishikawa, Yuichi, Takeuchi, Kengo
• Primary Examiner(s): Lee, Young
• Assistant Examiner(s): CARTER, RICHARD BRUCE
• Application Number:
  US12/950,268
• Publication Number:
  US 20110212486A1
• Time in Patent Office: 1,733 Days
• Field of Search: 348/79, 348/302, 356/338, 382/128, 382/133, 382/162, 435/29, 435/40.5
• US Class Current: 1/1
• 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: G02B 21/365 Control or image processing arrangements for digital or video microscopes G02B21/361, G02B21/362 take precedence

Abstract

A microscope system includes an acquisition unit that obtains a specimen image acquired by capturing the specimen stained by an element identification dye that visualizes a predetermined cell constituent element and a molecule target dye that visualizes a predetermined target molecule by using a microscope; a dye amount unit that obtains dye amounts of the element identification dye and the molecule target dye that stain corresponding positions on the specimen for each pixel of the image; an element area identification unit that identifies an area of the cell constituent element based on the dye amount of the element identification dye; a condition setting unit that sets the presence or absence of the predetermined target molecule on the cell constituent element as a condition; and an extraction unit that extracts an area of a target portion that satisfies the condition based on the dye amount of the molecule target dye.

Claims

1. A microscope system comprising:

an image acquisition unit that obtains, using a microscope, a specimen image acquired by capturing an image of the specimen stained by a plurality of cell element identification dyes that identifies two or more predetermined cell constituent elements based upon stain intensities produced by an interaction of the cell element identification dyes and the predetermined cell constituent elements, and a molecule target dye that identifies a predetermined target molecule based upon a stain intensity produced by an interaction of the molecule target dye and the predetermined target molecule;

a stain intensity obtaining unit that obtains stain intensities of the cell element identification dyes and the molecule target dye that stain corresponding positions on the specimen for each pixel of the acquired specimen image;

an element area identification unit that identifies areas of the two or more predetermined cell constituent elements within the specimen image on the basis of the obtained stain intensities of the cell element identification dyes;

a cell area recognition unit for recognizing an area of a cell determined by at least one of the areas of the two or more cell constituent elements;

a characteristic amount calculator that calculates a morphological characteristic amount of the cell;

an extraction condition setting unit that sets the presence or absence of the predetermined target molecule on the basis of the morphological characteristic amount of the cell and the obtained stain intensity of the molecule target dye, at least within the plurality of the identified area of the predetermined cell constituent elements as an extraction condition;

a target portion extraction unit that extracts an area of a target portion that satisfies the set extraction condition on the basis of identifying within at least one of the identified areas of the predetermined cell constituent element of the specimen image pixels that represent the presence of the stain intensities of the molecule target dye, if any, and considering the calculated morphological characteristic amount of the cell to which at least one of the two predetermined cell constituent elements belongs;

a display image generator that generates a display image representing the area of the target portion; and

a display processing unit that performs processing for displaying the display image on a display unit.

2. The microscope system according to claim 1, wherein

the extraction condition setting unit further sets a presence density of the predetermined target molecule within at least one of the identified areas of the predetermined cell constituent elements as the extraction condition, and

the target portion extraction unit extracts the area of the target portion based on the obtained stain intensity of the molecule target dye.

3. The microscope system according to claim 1, wherein

the extraction condition setting unit further sets a presence rate of the predetermined target molecule within at least one of the identified area of the predetermined cell constituent elements as the extraction condition, and

the target portion extraction unit extracts the area of the target portion based on the number of pixels that include the molecule target dye within at least one of the identified area of the predetermined cell constituent elements.

4. The microscope system according to claim 1, wherein

the specimen is stained by a plurality of molecule target dyes that identify two or more target molecules, and

the extraction condition setting unit sets the presence or absence of each of a plurality of target molecules within at least one of the identified area of the predetermined cell constituent elements as the extraction condition.

5. The microscope system according to claim 1, further comprising an abnormality degree determination unit for determining an abnormality degree of the cell on the basis of the morphological characteristic amount calculated by the characteristic amount calculator,

wherein the extraction condition setting unit further sets an abnormality degree of a cell as the extraction condition, and

the target portion extraction unit extracts the area of the target portion considering an abnormality degree of a cell to which at least one of the predetermined cell constituent elements belongs.

6. The microscope system according to claim 1, further comprising a statistics calculator for calculating at least one of the number of the predetermined cell constituent elements that include the area of the target portion extracted by the target portion extraction unit and a rate of the number of the predetermined cell constituent elements that include the area of the target portion to the total number of the identified predetermined cell constituent elements present in the specimen.

7. The microscope system according to claim 1, wherein the at least one of the predetermined cell constituent elements is a cell membrane, a cell nucleus, or a cytoplasm.

8. The microscope system according to claim 1, wherein the image acquisition unit acquires a plurality of specimen images by capturing images of portions of the specimen while relatively moving the specimen and an objective lens in a plane perpendicular to an optical axis of the objective lens, and

The microscope system further includes a specimen image generator for generating a single specimen image by combining the plurality of specimen images.

9. A specimen observation method comprising:

obtaining, using a microscope, a specimen image acquired by capturing an image of the specimen stained by a plurality of cell element identification dyes that identifies two or more predetermined cell constituent elements based upon stain intensities produced by an interaction of the cell element identification dyes and the predetermined cell constituent elements, and a molecule target dye that identifies a predetermined target molecule based upon a stain intensity produced by an interaction of the molecule target dye and the predetermined target molecule;

obtaining stain intensities of the cell element identification dyes and the molecule target dye that stain corresponding positions on the specimen for each pixel of the specimen image;

identifying area of the predetermined cell constituent elements in the specimen image on the basis of the obtained stain intensities of the cell element identification dyes;

recognizing an area of a cell determined by at least one of the areas of the two or more cell constituent elements;

calculating a morphological characteristic amount of the cell;

setting the presence or absence of the predetermined target molecule on the basis of the morphological characteristic amount of the cell and the obtained stain intensity of the molecule target dye, at least within the plurality of the identified areas of the predetermined cell constituent elements as an extraction condition;

extracting an area of a target portion that satisfies the extraction condition on the basis of identifying within the at least one identified area of the predetermined cell constituent elements of the specimen image pixels that represent the presence of the stain intensities of the molecule target dye, if any, and considering the calculated morphological characteristic amount of the cell to which at least one of the two predetermined cell constituent elements belongs;

generating a display image representing the area of the target portion; and

displaying the display image.

10. A computer program product having a non-transitory computer readable medium including programmed instructions, wherein the instructions, when executed by a computer, cause the computer to perform:

obtaining, using an operation instruction to a microscope, a specimen image acquired by capturing an image of the specimen stained by a plurality of cell element identification dyes that identifies two or more predetermined cell constituent elements based upon stain intensities produced by an interaction of the cell element identification dyes and the predetermined cell constituent elements, and a molecule target dye that identifies a predetermined target molecule based upon a stain intensity produced by an interaction of the molecule target dye and the predetermined target molecule;

obtaining stain intensities of the cell element identification dyes and the molecule target dye that stain corresponding positions on the specimen for each pixel of the specimen image;

identifying areas of the predetermined cell constituent elements in the specimen image on the basis of the obtained stain intensities of the cell element identification dyes;

recognizing an area of a cell determined by at least one of the areas of the two or more cell constituent elements;

calculating a morphological characteristic amount of the cell;

setting the presence or absence of the predetermined target molecule on the basis of the morphological characteristic amount of the cell and the obtained stain intensity of the molecule target dye, at least within the plurality of the identified areas of the predetermined cell constituent elements as an extraction condition;

extracting an area of a target portion that satisfies the extraction condition on the basis of identifying within the at least one identified areas of the predetermined cell constituent elements of the specimen image pixels that represent the presence of the stain intensities of the molecule target dye, if any, and considering the calculated morphological characteristic amount of the cell to which at least one of the two predetermined cell constituent elements belongs;

generating a display image representing the area of the target portion; and

displaying the display image.