Biological substance quantification method, image processing device, pathological diagnosis support system, and recording medium storing computer readable program

An image processing device that quantifies an expression amount of a specific biological substance in a target sample which is stained by using a fluorescent dye accumulated particle bondable to the specific biological substance, the device comprising:

• an inputter to input a first fluorescent image obtained by image capturing of the target sample;
• and
• a hardware processor that:
• extracts a bright spot portion from the first fluorescent image and calculates a first luminance integrated value which is an integrated value of a luminance value of the bright spot portion;
• and
• calculates a number of the fluorescent dye accumulated particle included in the bright spot portion extracted from the first fluorescent image by using the first luminance integrated value, a second luminance integrated value and a distribution of a third luminance integrated value, the second luminance integrated value being an integrated value of a luminance value of a bright spot portion extracted from a second fluorescent image obtained by image capturing of a standard sample for which an expression amount of the specific biological substance is measured in advance, and the third luminance integrated value adding up a luminance value of each bright spot portion representing light emission of the fluorescent dye accumulated particle in a third fluorescent image obtained by image capturing of a preparation on which the fluorescent dye accumulated particle is dispersed without aggregating;
• each of the bright spot portion is a region where the fluorescent dye accumulated particle exists, the first luminance value is a first luminance integrated value that is an integrated value of the luminance value of the bright spot portion extracted from the first fluorescent image, the second luminance value is a second luminance integrated value that is an integrated value of the luminance value of the bright spot portion extracted from the second fluorescent image, and the third luminance value is a third luminance integrated value that is an integrated value of the luminance value of the bright spot portion extracted from the third fluorescent image;
• a unit luminance value calculating comprising calculating a unit luminance value per the fluorescent dye accumulated particle from the distribution of the third luminance integrated value adding up the luminance value of each of the bright spot portion representing the light emission of the fluorescent dye accumulated particle in the third fluorescent image;
• a tentative particle number calculating comprising calculating a tentative number of the fluorescent dye accumulated particle included in the bright spot portion extracted from the first fluorescent image by using a first luminance integrated value and the unit luminance value;
• and
• a particle number correcting comprising correcting the tentative number of the fluorescent dye accumulated particle included in the bright spot portion extracted from the first fluorescent image by using the second luminance integrated value.

A non-transitory recording medium storing a computer readable program causing a computer that quantifies an expression amount of a specific biological substance in a target sample which is stained by using a fluorescent dye accumulated particle bondable to the specific biological substance to perform:

• inputting that is inputting a first fluorescent image obtained by image capturing of the target sample;
• luminance calculating that is extracting a bright spot portion from the first fluorescent image and calculating a first luminance integrated value which is an integrated value of a luminance value of the bright spot portion;
• and
• particle number calculating that is calculating a number of the fluorescent dye accumulated particle included in the bright spot portion extracted from the first fluorescent image by using the first luminance integrated value, a second luminance integrated value and a distribution of a third luminance integrated value, the second luminance integrated value being an integrated value of a luminance value of a bright spot portion extracted from a second fluorescent image obtained by image capturing of a standard sample for which an expression amount of the specific biological substance is measured in advance, and the third luminance integrated value adding up a luminance value of each bright spot portion representing light emission of the fluorescent dye accumulated particle in a third fluorescent image obtained by image capturing of a preparation on which the fluorescent dye accumulated particle is dispersed without aggregating;
• and
• each of the bright spot portion is a region where the fluorescent dye accumulated particle exists, the first luminance value is a first luminance integrated value that is an integrated value of the luminance value of the bright spot portion extracted from the first fluorescent image, the second luminance value is a second luminance integrated value that is an integrated value of the luminance value of the bright spot portion extracted from the second fluorescent image, and the third luminance value is a third luminance integrated value that is an integrated value of the luminance value of the bright spot portion extracted from the third fluorescent image;
• a unit luminance value calculating comprising calculating a unit luminance value per the fluorescent dye accumulated particle from the distribution of the third luminance integrated value adding up the luminance value of each of the bright spot portion representing the light emission of the fluorescent dye accumulated particle in the third fluorescent image;
• a tentative particle number calculating comprising calculating a tentative number of the fluorescent dye accumulated particle included in the bright spot portion extracted from the first fluorescent image by using a first luminance integrated value and the unit luminance value;
• and
• a particle number correcting comprising correcting the tentative number of the fluorescent dye accumulated particle included in the bright spot portion extracted from the first fluorescent image by using the second luminance integrated value.