Cell cycle-related changes in nuclear chromatin of stimulated lymphocytes as measured by flow cytometry.
作者： Zbigniew Darzynkiewicz , Thomas K. Sharpless , Frank Traganos , Myron R. Melamed
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摘要: Flow cytometric techniques have been developed to assay lymphocyte stimulation as reflected by the increase in cell transcriptional activity and progression through cycle. The metachromatic fluorescent dye, acridine orange, is used (a) stain DNA RNA differentially individual cells, (b) nuclear chromatin after removal of cellular BY RNase pretreatment at acidic pH. Stimulated cells with diploid content (G1) an increased stainable that makes it possible distinguish them from nonstimulated (G0) cells. G0 can also be distinguished G1 based on differences stainability their treatment acid. Mitotic indices scored automatically, inasmuch metaphase stains differently than does interphase Altogether, numbers G0, G1, S, G2, M phases may obtained rapidly great accuracy. transciptional correlated changes (e.g., during transition G1). two independent prove useful recognizing quantitating noncycling other systems. mechanisms responsible for differential different cycle are discussed.
参考文章(14)
Renato Baserga, Multiplication and division in mammalian cells ,(1976)
Peter C. Nowell, Phytohemagglutinin: an initiator of mitosis in cultures of normal human leukocytes. Cancer Research. ,vol. 20, pp. 462- 466 ,(1960)
F Traganos, Z Darzynkiewicz, T Sharpless, M R Melamed, Simultaneous staining of ribonucleic and deoxyribonucleic acids in unfixed cells using acridine orange in a flow cytofluorometric system. Journal of Histochemistry and Cytochemistry. ,vol. 25, pp. 46- 56 ,(1977) , 10.1177/25.1.64567
Juan A. Subirana, Studies on the thermal denaturation of nucleohistones. Journal of Molecular Biology. ,vol. 74, pp. 363- 386 ,(1973) , 10.1016/0022-2836(73)90378-1
F Traganos, A J Gorski, Z Darzynkiewicz, T Sharpless, M R Melamed, Rapid multiparameter analysis of cell stimulation in mixed lymphocyte culture reactions. Journal of Histochemistry and Cytochemistry. ,vol. 25, pp. 881- 887 ,(1977) , 10.1177/25.7.142787
Thomas Y. Shih, Robert S. Lake, Studies on the structure of metaphase and interphase chromatin of Chinese hamster cells by circular dichroism and thermal denaturation. Biochemistry. ,vol. 11, pp. 4811- 4817 ,(1972) , 10.1021/BI00775A026
R.R. Liedeman, N.P. Matveyeva, S.A. Vostricova, L.L. Prilipko, Extrinsic factors affecting the binding of acridine orange to the DNP complex of cell nuclei in different physiological states Experimental Cell Research. ,vol. 90, pp. 105- 110 ,(1975) , 10.1016/0014-4827(75)90362-6
Z. Darżynkiewicz, F. Traganos, T. Sharpless, M.R. Melamed, Conformation of RNA in situ as studied by acridine orange staining and automated cytofluorometry. Experimental Cell Research. ,vol. 95, pp. 143- 153 ,(1975) , 10.1016/0014-4827(75)90619-9
L. S. Lerman, The structure of the DNA-acridine complex. Proceedings of the National Academy of Sciences of the United States of America. ,vol. 49, pp. 94- 102 ,(1963) , 10.1073/PNAS.49.1.94
M. R. Melamed, F. Traganos, Z. Darzynkiewicz, T. Sharpless, Nucleic acid content and cell cycle distribution of five human bladder cell lines analysed by flow cytofluorometry International Journal of Cancer. ,vol. 20, pp. 30- 36 ,(1977) , 10.1002/IJC.2910200107