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Pairs of fluorescent dyes as probes of DNA and chromosomes

SA Latt, E Sahar and ME Eisenhard

If two fluorescent dyes with different binding or fluorescence specificities are used simultaneously to stain DNA or chromosomes, the ratio of their fluorescent signals can provide information about base composition or base analogue substitution. Energy transfer between such dye pairs, possible if the fluorescence spectrum of one overlaps the absorption spectrum of the other, can modify observed fluorescence. Microfluorometric measurements were used to document the occurrence of energy transfer between quinacrine or 33258 Hoechst as energy donor and ethidium or 7-aminoactinomycin D as acceptor when used jointly to stain cytologic preparations of human metaphase chromosomes. Use of 7- aminoactinomycin D, a dye with G-C binding specificity, as energy acceptor permitted the identification of human chromosome regions presumptively enriched for clusters of A-T base pairs, based on the resistance of A-T specific fluorescence, from quinacrine or 33258 Hoechst, to energy transfer dependent quenching. The results provide information about basic structural features of metaphase chromosomes, and the associated methodology may prove useful in accentuating specific fluorescent polymorphic chromosome regions.

Volume 27, Issue 1, pp. 65-71, 01/01/1979
Copyright © 1979 by The Histochemical Society


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S.-i. Murata, P. Herman, K. Mochizuki, T. Nakazawa, T. Kondo, N. Nakamura, J. R. Lakowicz, and R. Katoh
Spatial Distribution Analysis of AT- and GC-rich Regions in Nuclei Using Corrected Fluorescence Resonance Energy Transfer
J. Histochem. Cytochem., July 1, 2003; 51(7): 951 - 958.
[Abstract] [Full Text] [PDF]



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