New tools are desirable to examine the metabolic state of individual cells within tissues. We proposed a fluorescence-based procedure consisting of acridine orange staining and fast green counterstaining (AO-FG) to improve the selectivity of the former for nucleic acids (acridine orange stains both DNA and RNA with different fluorescence colors), with no interference from proteins. We compared this test with the biochemical quantification of the relative amounts of RNA and DNA in selected rat ventral prostate samples and PC3 cells. The epithelium of the prostate gland is highly active metabolically for the production of secretions. Differences in AO-RNA staining were revealed and correlated with the metabolic state of the epithelium. Specificity was confirmed by RNase A. To assess how AO-FG staining correlates with the metabolic state of the cell, we cultured PC3 cells in different concentrations of glucose and measured the ratios between the amounts of RNA and DNA. In parallel, similar cultures were subjected to AO-FG, and the staining pattern correlated closely (r2=0.886) with the obtained biochemical results. The results confirmed that the combined use of AO and FG is useful for detecting DNA and RNA simultaneously, as well as for assessing quantitatively the transcriptional activity of individual cells and their changes in response to experimental manipulation.

中文翻译：

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DNA / RNA的改良a啶橙染色。

需要新的工具来检查组织内单个细胞的代谢状态。我们提出了一种基于荧光的方法，该方法包括a啶橙染色和快速绿色复染（AO-FG），以提高前者对核酸的选择性（ac啶橙将DNA和RNA都用不同的荧光色染色），而不受蛋白质的干扰。我们将该测试与所选大鼠腹侧前列腺样品和PC3细胞中RNA和DNA相对量的生化定量进行了比较。前列腺的上皮在代谢上具有高活性，用于分泌物的产生。揭示了AO-RNA染色的差异，并与上皮的代谢状态相关。RNase A证实了特异性。为评估AO-FG染色如何与细胞的代谢状态相关，我们用不同浓度的葡萄糖培养PC3细胞，并测量RNA和DNA量之间的比率。同时，对相似的培养物进行AO-FG染色，染色模式与所获得的生化结果密切相关（r2 = 0.886）。结果证实，AO和FG的组合使用可用于同时检测DNA和RNA，以及定量评估单个细胞的转录活性及其响应实验操作的变化。