Abstract: An image processing using a plurality of specimen images obtained by successively applying a plurality of types of staining to a specimen to be evaluated and imaging the specimen after staining for at least two types of staining is performed. The image processing comprises: extracting an inner region corresponding to inward of a cell membrane of a single cell in the specimen based on at least one of the specimen images, specifying a cell region corresponding to an individual cell included in the specimen by expanding the inner region outwardly, and performing image cytometry for the cell region based on the specimen images. In analyzing a pathological specimen on a cell-by-cell basis, it is possible to deal with multiple immunostaining and specify the positions of individual cells and evaluate each cell separately even when a cell density in the specimen is high.

Abstract: Immunohistochemical (IHC) techniques that enable the sequential evaluation of at least seven biomarkers in one formalin-fixed paraffin-embedded (FFPE) tissue section are disclosed. The methods involve high-throughput multiplexed, quantitative IHC imaging, sequential IHC with iterative labeling, digital scanning, image coregistration and merging, and subsequent stripping of sections.