Open AccessPreparation of monolayer smears from paraffin‐embedded tissue for image cytometry
Author(s) -
van DrielKulker A. M. J.,
Mesker W. E.,
van Velzen I.,
Tanke H. J.,
Feichtinger J.,
Ploem J. S.
Publication year - 1985
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990060314
Subject(s) - monolayer , staining , feulgen stain , cytometry , population , pathology , flow cytometry , cell , biomedical engineering , chromatography , biology , microbiology and biotechnology , materials science , chemistry , medicine , nanotechnology , biochemistry , environmental health
A method is described for the preparation of monolayer smears from paraffin‐embedded tissue suitable for automated image analysis and DNA measurements. The proposed technique uses enzyme treatment and syringing for cell dispersal. Slide preparation is performed by centrifugal cytology. After Feulgen staining the quality of the monolayer smears is sufficiently high to enable visual morphologic evaluation. Automated DNA measurements using the Leyden television analysis system (LEYTAS) show coefficients of variation (CV) of 4.5% for the diploid cell population of the suspended tissue. This is approximately the same as the CV in fresh material from the same tumor. Formalin fixed trout red blood cells are used as reference cells. By applying image cytometry to paraffin‐embedded tissue this method allows retrospective studies of, for instance, the significan of DNA content with regard to the behavior of a tumor.