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 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 object of the present invention is to provide a non-clinical test method which allows for the quality control of experimental animals having a transplanted tumor site, such as tumor-bearing mice, or experimental animals having a lesion site other than a transplanted a tumor site, and which is characterized by including the step of identifying, using a specimen collected from such an experimental animal, the profile of a lesion site, for example, a transplanted tumor site, of the experimental animal, by a quantitative technique.

Abstract: The present invention relates to a method relating the evaluation of tumor tissue from an experimental animal, the method including the step of measuring an amount of a substance, except for human-derived tumor cells, contained in a sample collected from a region in an experimental animal into which a human-derived tumor tissue or tumor cells have been transplanted, wherein the region contains the tumor tissue or the tumor cells.

Abstract: For fluorescent nanoparticles having a zeta potential of ?10 mV to ?60 mV at pH 7.0 or a zeta potential of 0 mV to ?10 mV in a buffer of pH 6.0 to 8.0, an appropriate electrical repulsive force can be generated between biomolecules that are generally negatively charged and the fluorescent nanoparticles. As a result, non-specific binding between the fluorescent nanoparticles and the biomolecules is suppressed and the fluorescent nanoparticles are specifically bound to a biomolecule to be stained through interaction stronger than the electrical repulsive force, so that the visibility of the specific biomolecule to be stained can be improved. Further, since an appropriate electrical repulsive force is also generated between the fluorescent nanoparticles themselves, aggregation of the fluorescent nanoparticles can be inhibited and the dispersibility in a staining solution can thereby be maintained.

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: An analysis device for an objective biological substance includes a generator, a divider, an analyzer, and a calculator. The generator generates a microscopic image of a tissue sample after staining. The divider divides the microscopic image into at least one section having a prescribed size. The analyzer analyzes a staining condition of the microscopic image for each section. The calculator calculates a prescribed statistic based on an analysis result by the analyzer.

Abstract: Provided is a means for detecting and quantifying a biological substance of interest with an improved accuracy by inhibiting non-specific adsorption of fluorescent nanoparticles and thereby reducing the background noise in immunostaining with fluorescent nanoparticles. Immunostaining is carried out upon diluting fluorescent nanoparticles with a fluorescent nanoparticle diluent which contains 1 to 5% (W/W) of a protein having a molecular weight of 40,000 or higher (e.g., BSA) and 1 to 3% (W/W) of a protein having a molecular weight of less than 40,000 (e.g., casein) and, when casein is used as a low-molecular-weight protein, it is preferred that the ?-casein content in the casein is 10% (W/W) or less and the ratio of ?-casein and ?-casein (?-casein:?-casein) contained in the casein is 40:60 to 60:40 (taking the total amount of ?-casein and ?-casein as 100).

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: 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: Provided are: a phosphor-integrated nanoparticle labeling agent which is capable of yielding a sufficient signal intensity even when its final concentration in an immunofluorescent staining reaction system is low (e.g., 0.02 nM) and enables to obtain an immunofluorescently stained image with reduced noise by inhibiting non-specific adsorption of a probe biological substance and a label to substances other than a detection subject; and an immunostaining method using the same. The phosphor-integrated nanoparticle labeling agent is a set which includes: a probe biological substance 3, which is linked to a first binding group substance A via a first hydrophilic polymer-derived spacer 1 having a length of 30 ? to 1,000 ? and specifically binds to a biomolecule 2; and a phosphor-integrated nanoparticle 5, which has a second binding group substance B capable of specifically binding to the first binding group substance A.

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: 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: 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 object of the present invention is to provide: a staining agent for tissue staining which has an improved fluorescence signal evaluation accuracy; and a tissue staining kit comprising the staining agent. The staining agent for tissue staining contains, as a staining component, dye-resin particles comprising thermosetting resin particles and a fluorescent dye immobilized on the resin particles, wherein the resin particles contains a substituent having an electric charge opposite to that of the fluorescent dye and forms an ionic bond or a covalent bond with the fluorescent dye, and the dye-resin particles have a particle size variation coefficient of 15% or less.

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.