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Updated: May 15, 2026
Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
Immunohistochemical evaluation of cytoplasmically localized prostate-specific antigen expression in prostate cancer
Radoslaw Minias1, Krzysztof W Zielinski
1Department of Pathomorphology and Clinical Cytopathology, Medical University of Lodz, 113 Zeromskiego Street, 90-549 Lodz, Poland. radoslaw.minias@umed.lodz.pl
Objective:
To quantify the densitometric expression of prostate-specific antigen (PSA) in cytoplasm of normal prostate glandular cells and prostate cancer cells of various Gleason's pattern.
Study Design:
A total of 96 microphotographs showing neoplastic (48) and benign (48) prostatic tissues obtained from 48 patients were included in the study. The specimens were formalin-fixed, paraffin-embedded, stained with hematoxylin-eosin and immunostained with anti-PSA antibody and DAB-based detection system. The ImageJ software was used as an image analysis system. With this system the images of glandular tissue were divided into individual cells using the Voronoi tessellation method. The crude and standardized optical densities as well as the basic morphometric features of cells were measured.
Results:
There is a strong statistically significant correlation between the value of optical density of anti-PSA immunostained prostate cancer cells and the level of histological differentiation (Gleason's pattern). Gleason pattern 5 cell optical density histogram was bimodal, which means that there are 2 cell populations which are statistically significantly different.
Conclusion:
The Voronoi tessellation is an effective method for dividing the immunostained images with cytoplasmically-located antigens into single cells.
