A highly accurate and quantitative method for determining
cancer onset or
cancer onset risk by a quantitative
tissue staining method in biological tissues using an
antibody capable of recognizing a
cancer growth regulatory factor or
cancer metastasis regulatory factor such as PAR1
antibody, which inhibits the
cancer cell mobility and infiltration is provided.
Cancer onset or cancer onset risk is determined using the
tissue staining method comprising the steps of: labeling an
antibody which recognizes a cancer growth regulatory factor or
cancer metastasis regulatory factor with a fluorescent material, and contacting the fluorescent-labeled antibody with a
tissue sample; irradiating a tissue site in contact with the antibody with excitation light to acquire a
fluorescence image; acquiring an
autofluorescence image in a vicinity region of a short
wavelength side or
long wavelength side of an acquisition region of
fluorescence wavelength emitted by the fluorescent material, in the same field of vision and in the same focal point as those of the
fluorescence image; acquiring a corrected fluorescence image by
image processing to eliminate a fluorescent brightness of the
autofluorescence image from the fluorescent brightness of the fluorescence image; counting the number of cells at the tissue site in contact with the antibody; measuring a mean fluorescent brightness of a single
fluorescent particle; and calculating the number of fluorescent particles per
cell.