Latt S A, Sahar E, Eisenhard M E
J Histochem Cytochem. 1979 Jan;27(1):65-71. doi: 10.1177/27.1.86582.
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.
如果同时使用两种具有不同结合或荧光特异性的荧光染料对DNA或染色体进行染色，它们荧光信号的比率可以提供有关碱基组成或碱基类似物取代的信息。如果一种染料的荧光光谱与另一种染料的吸收光谱重叠，那么这种染料对之间就可能发生能量转移，从而改变观察到的荧光。当喹吖因或33258 Hoechst作为能量供体与溴化乙锭或7-氨基放线菌素D作为受体联合用于染色人类中期染色体的细胞学制剂时，利用显微荧光测量记录了能量转移的发生情况。使用具有G-C结合特异性的染料7-氨基放线菌素D作为能量受体，基于来自喹吖因或33258 Hoechst的A-T特异性荧光对能量转移依赖性猝灭的抗性，能够鉴定出推测富含A-T碱基对簇的人类染色体区域。这些结果提供了有关中期染色体基本结构特征的信息，并且相关方法可能在突出特定荧光多态性染色体区域方面证明是有用的。
