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: 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 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 detection method for detecting a biological substance specifically in a pathological specimen, comprising 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: Provided is a method for staining a tissue enabling highly precise staining, by which the expression amount and/or the location of a biological substance in a tissue sample can be detected with a high quantitativity together with detailed information that can be obtained by bright field observation. The tissue staining method of the present invention is a method for staining a tissue, in which both staining that allows bright field observation and fluorescence staining are carried out for the same specific biological substance.

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: 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 method of producing core/shell-type fluorescent dye-containing nanoparticles for immunohistochemical staining or live cell imaging, the method including: the step 1 of polymerizing monomers for thermoplastic resin synthesis in the presence of a fluorescent dye and thereby preparing core particles composed of a thermoplastic resin containing the fluorescent dye; and the step 2 of coating the core particles each with a shell layer composed of a thermosetting resin. By the method of producing fluorescent dye-containing nanoparticles according to the present invention, fluorescent dye-containing nanoparticles having a high brightness, whose dye does not elutes into water, physiological saline, culture medium and the like, can be produced, and the fluorescent dye-containing nanoparticles can be effectively utilized in immunohistochemical staining and live cell imaging.

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: [Problem ] 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. [Means for Solution] 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 (eg ., 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: 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: A detection method for a specific biological substance uses, as a color former, fluorescent substance-encapsulated nanoparticles which have biological substance-recognizing molecules. The biological substance-recognizing molecules specifically recognize a specific biological substance. The biological substance-recognizing molecules are bonded to the surface of the nanoparticles. Nanoparticles encapsulating no fluorescent substance are used as a blocking agent for preventing the fluorescent substance-encapsulated nanoparticles from being non-specifically adsorbed on a biological substance other than the specific biological substance.

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 surppressed 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: The present invention provides a fluorescent label that can be used for carrying out 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 fluorescent label is a fluorescent dye-containing nanoparticle in which the parent material is a cross-linked polymer and the fluorescent dye is an aromatic ring-based dye molecule. The cross-linked polymer is suitably a melamine resin or a styrene resin. The aromatic ring-based dye molecule is suitably a perylene and more suitably a perylene diimide.

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: An object of the present invention is to provide a fluorophore having excellent initial emission intensity and light resistance, which is not easily discolored and does not affect the evaluation of the number of bright spots even when it is slightly discolored. The resin particle for fluorescence labeling is characterized by comprising a fluorescent dye immobilized in a resin particle, the fluorescent dye satisfying both of the following conditions (1) and (2): (1) the fluorescent dye alone in an aqueous solution has a concentration-dependent peak emission intensity in a range of 30 to 80 ?M; and (2) the emission intensity at a concentration of 100 ?M is not less than 80% of the peak emission intensity. It is preferred that the Stoke's shift of the fluorescent dye in an aqueous solution is not less than 25 nm. The fluorescent dye may be subjected to a solubilization treatment (such as an acid treatment).

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: Provided is a method for staining a tissue enabling highly precise staining, by which the expression amount and/or the location of a biological substance in a tissue sample can be detected with a high quantitativity together with detailed information that can be obtained by bright field observation. The tissue staining method of the present invention is a method for staining a tissue, in which both staining that allows bright field observation and fluorescence staining are carried out for the same specific biological substance.

Abstract: It is an object of the present invention to provide a detection method in which background noise is not increased even if a high-luminance fluorescent labeling material is used and which has an improved S/N ratio and high quantitativity and is advantageous for an immunohistological staining method.