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Acid giemsa technique for rapid identification of mitotic cells

WC Dooley, J Roberts and DC Allison

Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.

We developed a rapid technique for differential staining of compacted chromatin as a tool for screening of large tissue culture cell populations for mitotic cells. With a combination of acid Giemsa staining and counterstaining, differential staining of mitotic cells and classification according to stage of mitosis can be accomplished at magnifications as low as x 50-100 (objectives of x 5-10). The mapped and classified cells can then be de-stained and re-studied for DNA content by Feulgen staining and/or for uptake of radioactive DNA precursors by autoradiography. The staining and de-staining procedures outlined do not affect the reproducibility and accuracy of DNA content measurements or measurements of radioactive uptake. Therefore, this technique can be used for cell kinetic analysis by the percentage labeled mitoses method and for cytophotometric studies of mitotic segregation.

Volume 37, Issue 10, pp. 1553-1556, 10/01/1989
Copyright © 1989 by The Histochemical Society


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