Abstract: The present invention provides: an aminocoumarin compound having a structure represented by the following formula (1); and an aminocoumarin compound-containing resin particle containing the aminocoumarin compound and a resin particle containing the aminocoumarin compound. In formula (1), Rs each independently represent a hydrogen atom or a methyl group, Q represents a sulfur atom, an oxygen atom, or N—R1, and R1 represents a hydrogen atom or a methyl group. The aminocoumarin compound of the present invention is a fluorescent dye that has a longer excitation wavelength than a conventionally known sulfonated coumarin-based compound and is effectively excited at a wavelength of 475 nm or more, particularly at a wavelength around 500 nm.

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 method of producing dye-containing thermosetting resin particles, the method including: dispersing seed particles formed from a thermosetting resin in a dispersion medium; and polymerizing a monomer for thermosetting resin synthesis in the thus obtained dispersion in the presence of a dye and an acid catalyst. By the method of producing dye-containing thermosetting resin particles according to the present invention, dye-containing thermosetting resin particles whose particle size variation coefficient is small at, for example, 8% or less, can be produced without performing a post-treatment such as centrifugation. Therefore, when immunological observation is performed by a fluorescent labeling method using fluorescent dye-containing resin particles produced by the method of producing dye-containing thermosetting resin particles according to the present invention, a high bright spot detection accuracy can be attained.

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: The present invention provides: an aminocoumarin compound having a structure represented by the following formula (1); and an aminocoumarin compound-containing resin particle containing the aminocoumarin compound and a resin particle containing the aminocoumarin compound. In formula (1), Rs each independently represent a hydrogen atom or a methyl group, Q represents a sulfur atom, an oxygen atom, or N—R1, and R1 represents a hydrogen atom or a methyl group. The aminocoumarin compound of the present invention is a fluorescent dye that has a longer excitation wavelength than a conventionally known sulfonated coumarin-based compound and is effectively excited at a wavelength of 475 nm or more, particularly at a wavelength around 500 nm.

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: The present invention provides a nanoparticle containing a hydrophobic fluorescent substance accumulated therein and a thermosetting resin as a matrix, which nanoparticle, when used for labeling of a biological substance such as a protein or nucleic acid, has brightness sufficient for allowing pathological diagnosis using a fluorescence image obtained thereby. The present invention is a phosphor integrated dots nanoparticles wherein a thermosetting resin contains a structural unit formed from a raw material containing a hydrophobic substituent, and wherein a fluorescent substance is accumulated in the nanoparticle at least by hydrophobic interaction, preferably further by stacking interaction, with the hydrophobic substituent of the thermosetting resin.

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 method of quantifying a specific biological material in a specimen stained using fluorescent dye accumulating particles capable of binding to the material includes: inputting a first fluorescence image obtained by capturing an image of the specimen; extracting a certain region from the first fluorescence image and calculating a first luminance integrated value by integrating luminance values of the certain region; and calculating the number of the particles included in the certain region from the first luminance integrated value and an average luminance value per fluorescent dye accumulating particle, wherein the average luminance value is calculated from a distribution of second luminance integrated values obtained by integrating luminance values for individual bright spot regions, which indicate emission of light by the fluorescent dye accumulating particles, in a second fluorescence image obtained by capturing an image of a preparation onto which the fluorescent dye accumulating particles are dispersed w

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: The present invention provides a method for evaluating a surface state of particles for judging suitability of colored particles used for staining a biological material. The method includes: a dispersion step of dispersing colored particles in a two-component dispersion medium containing two types of dispersion media having different polarities to form an interface; an indicator acquisition step of acquiring an indicator indicating the amount of particles contained in one of the two types of dispersion media; and a judgement step of judging, on the basis of the indicator, suitability of a surface state of the colored particles for staining a biological material.

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 method of quantifying a specific biological material in a specimen stained using fluorescent dye accumulating particles capable of binding to the material includes: inputting a first fluorescence image obtained by capturing an image of the specimen; extracting a certain region from the first fluorescence image and calculating a first luminance integrated value by integrating luminance values of the certain region; and calculating the number of the particles included in the certain region from the first luminance integrated value and an average luminance value per fluorescent dye accumulating particle, wherein the average luminance value is calculated from a distribution of second luminance integrated values obtained by integrating luminance values for individual bright spot regions, which indicate emission of light by the fluorescent dye accumulating particles, in a second fluorescence image obtained by capturing an image of a preparation onto which the fluorescent dye accumulating particles are dispersed w

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 method of producing dye-containing thermosetting resin particles, the method including: dispersing seed particles formed from a thermosetting resin in a dispersion medium; and polymerizing a monomer for thermosetting resin synthesis in the thus obtained dispersion in the presence of a dye and an acid catalyst. By the method of producing dye-containing thermosetting resin particles according to the present invention, dye-containing thermosetting resin particles whose particle size variation coefficient is small at, for example, 8% or less, can be produced without performing a post-treatment such as centrifugation. Therefore, when immunological observation is performed by a fluorescent labeling method using fluorescent dye-containing resin particles produced by the method of producing dye-containing thermosetting resin particles according to the present invention, a high bright spot detection accuracy can be attained.

Abstract: The present invention provides a nanoparticle containing a hydrophobic fluorescent substance accumulated therein and a thermosetting resin as a matrix, which nanoparticle, when used for labeling of a biological substance such as a protein or nucleic acid, has brightness sufficient for allowing pathological diagnosis using a fluorescence image obtained thereby. The present invention is a phosphor integrated dots nanoparticles wherein a thermosetting resin contains a structural unit formed from a raw material containing a hydrophobic substituent, and wherein a fluorescent substance is accumulated in the nanoparticle at least by hydrophobic interaction, preferably further by stacking interaction, with the hydrophobic substituent of the thermosetting resin.

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.