Simmons D M, Mercer A V, Hallas G, Dyson J E
University Department of Radiotherapy, Cookridge Hospital Leeds, United Kingdom.
J Histochem Cytochem. 1990 Jan;38(1):41-9. doi: 10.1177/38.1.1688448.
Few fluorescent stains specific for cell constituents other than DNA are available. To assess their potential use as fluorescent stains for flow cytometry, the cell staining specificity of 55 compounds, originally synthesized for use as textile dyes and fluorescent brighteners, was explored and their excitation and emission wavebands determined. From these, six dyes were chosen for more detailed analysis. All six are vital stains, with excitation wavelengths allowing their use with an argon ion laser, and specific for a range of cell structures including mitochondria, Golgi bodies, lipid droplets, nuclear membrane, and endoplasmic reticulum. Concentrations as low as 0.01-0.25 microM were found to be adequate for most purposes, and high background fluorescence was not a problem. Their specificity allows differentiation between non-cycling and cycling cells. The properties of two of the stains allows their combination with propidium iodide or ethidium bromide for simultaneous determination of DNA content profiles. Being vital stains, usable at very low concentrations, and specific for a range of cell organelles, these six stains may be of considerable utility in flow cytofluorometry. We suggest that other textile dyes may be of use in flow cytofluorometry, or that their structures may form a starting point for the synthesis of further fluorescent stains of enhanced specificity.
除了DNA之外，很少有针对细胞成分的特异性荧光染料。为了评估它们作为流式细胞术荧光染料的潜在用途，研究了最初作为纺织染料和荧光增白剂合成的55种化合物的细胞染色特异性，并确定了它们的激发和发射波段。从中选择了六种染料进行更详细的分析。所有六种都是活细胞染料，其激发波长允许它们与氩离子激光一起使用，并且对一系列细胞结构具有特异性，包括线粒体、高尔基体、脂滴、核膜和内质网。发现低至0.01 - 0.25 microM的浓度对于大多数目的来说就足够了，并且高背景荧光不是问题。它们的特异性允许区分非循环细胞和循环细胞。其中两种染料的特性使其能够与碘化丙啶或溴化乙锭结合，用于同时测定DNA含量分布。作为活细胞染料，可在非常低的浓度下使用，并且对一系列细胞器具有特异性，这六种染料在流式细胞荧光测定法中可能具有相当大的用途。我们认为其他纺织染料可能在流式细胞荧光测定法中有用，或者它们的结构可能成为合成具有更高特异性的进一步荧光染料的起点。
