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Chapter 13 Rapid DNA Content Analysis by the Propidium Iodide–Hypotonic Citrate Method

1990; Elsevier BV; Linguagem: Inglês

10.1016/s0091-679x(08)60518-x

0091-679X

Awtar Krishan,

Molecular Biology Techniques and Applications

For the rapid quantitation of DNA and protein content of fixed cells by flow cytometry (FCM), direct staining of isolated nuclei is experimented with various DNA-binding fluorochromes. The propidium iodide (PI)-citrate method has been developed for the rapid determination of cell cycle distribution in leukemic peripheral blood and bone marrow samples of patients on high-dose methotrexate rescue protocols. The method is employed and found suitable for rapid cell cycle analysis. It has also been used by several workers to study the cell cycle phase distribution of mammalian tissue culture cells and a variety of tumor cells. Modifications have involved inclusion of RNase and a detergent in the staining solution. This rapid method is, especially suitable for small samples, such as a drop of blood from a fingerprick, tissue culture cells from multiwell plates, and small bone marrow aspirates. For solid-tumor material, mincing of tissue with scissors or crossed scalpels and filtering through nylon cloth are essential.