Abstract: It is an object of the present invention to provide a tissue staining method that makes it possible to observe both information on the morphology of a tissue and information on a biological substance such as an antigen molecule to be detected on a single section and in a single view field. The present invention provides a tissue staining method, including carrying out (A) a HE (hematoxylin-eosin) staining, and (B) a histochemical staining, serially on a single tissue section, wherein the histochemical staining is defined as a histochemical technique for detecting a biological substance to be detected in a tissue in a visible manner by use of a binding reaction between the biological substance to be detected and a probe biological substance capable of binding specifically to the biological substance to be detected.

Abstract: A biological substance detection method for detecting a biological substance specifically in a pathological specimen, includes a step of immunologically staining the pathological specimen using a fluorescent label, a step of staining the pathological specimen with a staining reagent for morphology observation purposes (eosin) to observe the morphology of the pathological specimen, a step of irradiating the stained pathological specimen with excited light to cause the emission of a fluorescent and detecting the biological substance in the pathological specimen. In the step of immunologically staining the pathological specimen, a special fluorescent particle for which the excitation wavelength appears in a region that is different from the excitation wavelength region of eosin is used as the fluorescent label.

Abstract: The present invention relates to a test support method which is a means for predicting whether or not pathological complete response (pCR) will be attained when a preoperative chemotherapy with an anticancer drug is performed, the method using a sample (breast cancer tissue section) collected from a breast cancer patient to be subjected to the preoperative chemotherapy. The test support method includes: [1] the step of acquiring a fluorescence image of a breast cancer tissue section, which fluorescence image shows bright spots of fluorescent nanoparticles labeling one or more kinds of breast cancer-related proteins; [2] the step of acquiring at least one index relating to the expression level(s) of the breast cancer-related protein(s) on the basis of the bright spots of the fluorescence image; and [3] the step of acquiring information for predicting pCR by performing an analysis using the above-described at least one index.

Abstract: A tissue staining method which comprises: staining a tissue with a staining reagent wherein a biosubstance recognition site is bonded to particles carrying multiple fluorescent substances accumulated therein; in the stained tissue, counting fluorescent points or measuring fluorescent brightness; and evaluating the expression level of a biosubstance, which matches the biosubstance recognition site, in the aforesaid tissue on the basis of the number of the fluorescent points or fluorescent brightness that was measured.

Abstract: An image processing device (2A) comprises: an input means for inputting a brightfield image representing cell morphology in a tissue section, and a fluorescence image representing, by fluorescent bright spots, the expression of a specific protein in the same range of the tissue section; a first generation means for generating a cell image obtained by extracting a specific site of a cell from the brightfield image; a second generation means for generating an image obtained by extracting bright spot regions from the fluorescence image, creating a brightness profile for each bright spot region, and generating a fluorescent particle image obtained by extracting the fluorescent particles in the bright spot regions on the basis of the fluorescence profile for one fluorescent particle, which serves as a fluorescence bright spot source; and a calculation means for superimposing the cell image and the fluorescent particle image on one another.

Abstract: A tissue staining method which comprises: staining a tissue with a staining reagent wherein a biosubstance recognition site is bonded to particles carrying multiple fluorescent substances accumulated therein; in the stained tissue, counting fluorescent points or measuring fluorescent brightness; and evaluating the expression level of a biosubstance, which matches the biosubstance recognition site, in the aforesaid tissue on the basis of the number of the fluorescent points or fluorescent brightness that was measured.

Abstract: A tissue staining method which comprises: staining a tissue with a staining reagent wherein a biosubstance recognition site is bonded to particles carrying multiple fluorescent substances accumulated therein; in the stained tissue, counting fluorescent points or measuring fluorescent brightness; and evaluating the expression level of a biosubstance, which matches the biosubstance recognition site, in the aforesaid tissue on the basis of the number of the fluorescent points or fluorescent brightness that was measured.

Abstract: There is provided a biological substance quantitation method of quantitating a biological substance in a sample stained with a staining reagent including a fluorescent particle encapsulating a fluorescent substance, based on a fluorescence of the fluorescent substance. The method includes inputting a fluorescent image representing expression of the biological substance in the sample by a fluorescent bright spot; and quantitating an expression amount of the biological substance based on a fluorescence of the fluorescent bright spot. The biological substance is a nucleoprotein expressed at a cell nucleus. The fluorescent particle binds to the biological substance through a primary antibody which is directed against the biological substance as an antigen.

Abstract: It is an object of the present invention to provide a tissue staining method that makes it possible to observe both information on the morphology of a tissue and information on a biological substance such as an antigen molecule to be detected on a single section and in a single view field. The present invention provides a tissue staining method, including carrying out (A) a HE (hematoxylin-eosin) staining, and (B) a histochemical staining, serially on a single tissue section, wherein the histochemical staining is defined as a histochemical technique for detecting a biological substance to be detected in a tissue in a visible manner by use of a binding reaction between the biological substance to be detected and a probe biological substance capable of binding specifically to the biological substance to be detected.

Abstract: It is an object of the present invention to provide a tissue staining method that makes it possible to observe both information on the morphology of a tissue and information on a biological substance such as an antigen molecule to be detected on a single section and in a single view field. The present invention provides a tissue staining method, including carrying out (A) a HE (hematoxylin-eosin) staining, and (B) a histochemical staining, serially on a single tissue section, wherein the histochemical staining is defined as a histochemical technique for detecting a biological substance to be detected in a tissue in a visible manner by use of a binding reaction between the biological substance to be detected and a probe biological substance capable of binding specifically to the biological substance to be detected.

Abstract: The present invention provides a biological substance detection method for specifically detecting a biological substance from a pathological specimen, by which method, when immunostaining using a fluorescent label and staining for morphological observation using a staining agent for morphological observation are simultaneously performed, the results of fluorescence observation and immunostaining can be assessed properly even if the fluorescent label and/or the staining agent is/are deteriorated by irradiation with an excitation light. The biological substance detection method according to the present invention is characterized in that the brightness retention rate of an immunostained part is in a range of 80% to 120% in relation to the brightness retention rate of a part stained for morphological observation when the fluorescent label used for the immunostaining is observed.

Abstract: An image processing device (2A) comprises: an input means for inputting a brightfield image representing cell morphology in a tissue section, and a fluorescence image representing, by fluorescent bright spots, the expression of a specific protein in the same range of the tissue section; a first generation means for generating a cell image obtained by extracting a specific site of a cell from the brightfield image; a second generation means for generating an image obtained by extracting bright spot regions from the fluorescence image, creating a brightness profile for each bright spot region, and generating a fluorescent particle image obtained by extracting the fluorescent particles in the bright spot regions on the basis of the fluorescence profile for one fluorescent particle, which serves as a fluorescence bright spot source; and a calculation means for superimposing the cell image and the fluorescent particle image on one another.

Abstract: An image processing device (2A) comprises: an input means for inputting a brightfield image representing cell morphology in a tissue section, and a fluorescence image representing, by fluorescent bright spots, the expression of a specific protein in the same range of the tissue section; a first generation means for generating a cell image obtained by extracting a specific site of a cell from the brightfield image; a second generation means for generating an image obtained by extracting bright spot regions from the fluorescence image, creating a brightness profile for each bright spot region, and generating a fluorescent particle image obtained by extracting the fluorescent particles in the bright spot regions on the basis of the fluorescence profile for one fluorescent particle, which serves as a fluorescence bright spot source; and a calculation means for superimposing the cell image and the fluorescent particle image on one another.

Abstract: The present invention provides a staining method in which the fluorescent staining properties in a fluorescently-immunostained specimen are not reduced even when an oil-based mounting medium is used. The present invention also provides a method of preventing deterioration of a fluorescent label caused by irradiation with excitation light and improving the light resistance in a fluorescently-immunostained specimen obtained by the staining method. The biological substance detection method according to the present invention is a biological substance detection method for specifically detecting a biological substance from a pathological specimen, which includes the steps of: immunostaining the specimen with a fluorescent label; immobilizing the thus stained specimen; and mounting the thus immobilized specimen using a mounting medium including an organic solvent not freely miscible with water.

Abstract: An object of the present invention is to provide a method of detecting a specific tissue or cell in a sample tissue section and accurately specifying both the position(s) and amount of a biological substance of interest that is expressed on the specific tissue or cell.

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 florescent bright spot. A quantitative evaluation value of the florescent 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.

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 florescent bright spot. A quantitative evaluation value of the florescent 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.

Abstract: An image processing device (2A) comprises: an input means for inputting a brightfield image representing cell morphology in a tissue section, and a fluorescence image representing, by fluorescent bright spots, the expression of a specific protein in the same range of the tissue section; a first generation means for generating a cell image obtained by extracting a specific site of a cell from the brightfield image; a second generation means for generating an image obtained by extracting bright spot regions from the fluorescence image, creating a brightness profile for each bright spot region, and generating a fluorescent particle image obtained by extracting the fluorescent particles in the bright spot regions on the basis of the fluorescence profile for one fluorescent particle, which serves as a fluorescence bright spot source; and a calculation means for superimposing the cell image and the fluorescent particle image on one another.

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.

Abstract: An image processing device (2A) comprises: an input means for inputting a brightfield image representing cell morphology in a tissue section, and a fluorescence image representing, by fluorescent bright spots, the expression of a specific protein in the same range of the tissue section; a first generation means for generating a cell image obtained by extracting a specific site of a cell from the brightfield image; a second generation means for generating an image obtained by extracting bright spot regions from the fluorescence image, creating a brightness profile for each bright spot region, and generating a fluorescent particle image obtained by extracting the fluorescent particles in the bright spot regions on the basis of the fluorescence profile for one fluorescent particle, which serves as a fluorescence bright spot source; and a calculation means for superimposing the cell image and the fluorescent particle image on one another.