BIOLOGICAL SUBSTANCE QUANTITATION METHOD, PATHOLOGICAL DIAGNOSIS SUPPORT SYSTEM AND RECORDING MEDIUM STORING COMPUTER READABLE PROGRAM

• US 20180024059A1
• Filed: 02/10/2015
• Published: 01/25/2018
• Est. Priority Date: 02/10/2015
• Status: Active Grant

• Current Assignee: Konica Minolta Inc.
• Original Assignee: Konica Minolta Inc.
• Inventors: GOUDA, Hideki, TAKANASHI, Kensaku, GONDA, Kohsuke, OHUCHI, Noriaki, WATANABE, Mika
• Granted Patent: US 10,234,391 B2
• Time in Patent Office: Days
• 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 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 21/93 Detection standards; Calibrating baseline adjustment, drift correction
• CPC Class Codes: G01N 33/48 Biological material, e.g. blood, urine G01N33/02, G01N33/26, G01N33/44, G01N33/46 take precedence; Haemocytometers counting blood corpuscules distributed over a surface by scanning the surface G06M11/02
• CPC Class Codes: G01N 33/483 Physical analysis of biological material
• CPC Class Codes: G01N 33/5023 on expression patterns
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G06T 2207/10024 Color image
• CPC Class Codes: G06T 2207/10056 Microscopic image
• CPC Class Codes: G06T 2207/10064 Fluorescence image
• CPC Class Codes: G06T 2207/30024 Cell structures in vitro; Tissue sections in vitro
• CPC Class Codes: G06T 2207/30242 Counting objects in image
• CPC Class Codes: G06T 7/0012 Biomedical image inspection
• CPC Class Codes: G06T 7/0016 involving temporal comparison
• CPC Class Codes: G06T 7/90 Determination of colour characteristics

Abstract

A biological substance quantitation method includes the following. A fluorescent image is input, which represents an expression of a specific biological substance in a sample stained with a fluorescent substance by a fluorescent bright spot. A quantitative evaluation value of the fluorescent bright spot is calculated. A standard fluorescent image of a standard sample stained under a same condition as the sample and representing an expression of the biological substance in a standard sample, is input under a same condition as the fluorescent image. A quantitative evaluation value in the standard fluorescent image is calculated under a same condition as the fluorescent image. Based on a correlation between an expression amount of the biological substance in a standard sample measured in advance and the evaluation value in the standard fluorescent image, the evaluation value in the fluorescent image is converted to an expression amount of the biological substance in the sample.

Claims

1. A biological substance quantitation method which quantitates an expression amount of a specific biological substance in a sample stained with a staining reagent which stains the biological substance with a fluorescent substance, the method comprising:

inputting a fluorescent image which represents an expression of the biological substance in the sample by a fluorescent bright spot;

performing fluorescence quantitation which includes calculation of an evaluation value by quantitative evaluation of the fluorescent bright spot in the fluorescent image;

inputting a standard fluorescent image under a same condition as the inputting of the fluorescent image, wherein

the standard fluorescent image represents an expression of the biological substance in a standard sample by a fluorescent bright spot based on staining under a same condition as the sample, and

an expression amount of the biological substance in the standard sample is measured in advance;

performing standard fluorescence quantitation which includes calculation of an evaluation value by quantitative evaluation of the fluorescent bright spot in the standard fluorescent image under a same condition as the quantitation of fluorescence;

calculating a correlation between the expression amount of the biological substance in the standard sample and the evaluation value of the fluorescent bright spot in the standard fluorescent image; and

converting the evaluation value of the fluorescent bright spot in the fluorescent image to an expression amount of the biological substance in the sample based on the calculating of the correlation.

2. The biological substance quantitation method according to claim 1, wherein

a calibration curve is prepared in calculating the correlation, wherein the calibration curve represents the evaluation value of the fluorescent bright spot of the standard fluorescence image corresponding to the expression amount of the biological substance in the standard sample, and

in the converting, the evaluation value of the fluorescent bright spot of the fluorescent image is converted to the expression amount of the biological substance in the sample based on the calibration curve.

3. The biological substance quantitation method according to claim 1, wherein the standard sample is a cell cultured on a substrate.

4. The biological substance quantitation method according to claim 1, wherein the biological substance is a protein.

5. The biological substance quantitation method according to claim 1, wherein the staining reagent comprises a fluorescent particle in which a plurality of molecules of the fluorescent substance are accumulated.

6. A pathological diagnosis support system which quantitates an expression amount of a specific biological substance in a sample stained with a staining reagent which stains the biological substance with a fluorescent substance, the system comprising:

a fluorescent image inputting unit which inputs a fluorescent image which represents an expression of the biological substance in the sample by a fluorescent bright spot;

a fluorescence quantitation unit which calculates an evaluation value by quantitative evaluation of the fluorescent bright spot in the fluorescent image;

a standard fluorescent image inputting unit which inputs a standard fluorescent image under a same condition as the fluorescent image inputting unit, wherein

the standard fluorescent image represents an expression of the biological substance in a standard sample by a fluorescent bright spot based on staining under a same condition as the sample, and

an expression amount of the biological substance in the standard sample is measured in advance;

a standard fluorescent quantitation unit which calculates an evaluation value by quantitative evaluation of the fluorescent bright spot in the standard fluorescent image under a same condition as the fluorescence quantitation unit;

a correlation calculator which calculates a correlation between the expression amount of the biological substance in the standard sample and the evaluation value of the fluorescent bright spot in the standard fluorescent image; and

a converter which converts the evaluation value of the fluorescent bright spot in the fluorescent image to an expression amount of the biological substance in the sample based on the correlation.

7. A non-transitory recording medium storing a computer readable program causing a computer which quantitates an expression amount of a specific biological substance in a sample stained with a staining reagent which stains the biological substance with fluorescent substances, to function as:

a fluorescent image inputting unit which inputs a fluorescent image which represents an expression of the biological substance in the sample by a fluorescent bright spot;

a fluorescence quantitation unit which calculates an evaluation value by quantitative evaluation of the fluorescent bright spot in the fluorescent image;

a standard fluorescent image inputting unit which inputs a standard fluorescent image under a same condition as in the fluorescent image inputting unit, wherein

the standard fluorescent image represents an expression of the biological substance in a standard sample by a fluorescent bright spot based on staining under a same condition as the sample, and

an expression amount of the biological substance in the standard sample is measured in advance;

a standard fluorescent quantitation unit which calculates an evaluation value by quantitative evaluation of the fluorescent bright spot in the standard fluorescent image under a same condition as the fluorescence quantitation unit;

a correlation calculator which calculates a correlation between the expression amount of the biological substance in the standard sample and the evaluation value of the fluorescent bright spot in the standard fluorescent image; and

a converter which converts the evaluation value of the fluorescent bright spot in the fluorescent image to an expression amount of the biological substance in the sample based on the correlation.