Alvarado-Aleman F J, Collazo-Ponce A, Kumate-Rodríguez J
Departamento de Inmunoquímica Dr. Maximiliano Ruiz Castañeda, Instituto Nacional de Diagnóstico y Referencia Epidemiológicos, Dirección General de Epidemiología, SSA, Mexico.
Rev Latinoam Microbiol. 1996 Jan-Mar;38(1):25-30.
A comparative study of two different staining methods for estimation of yeast's viability is assessed by microbial flow cytometry. Propidium iodide (PI) a well known nucleic-acid binding vital dye exclusion is compared with light green (LG), a cytoplasmic function related vital dye widely used in histological, histochemical and immunocytochemical staining. The yeast model used was heat-killed Candida guilliermondii cells. The stained heat-killed bacterial population showed on the dot-plot flow cytometer analysis, a clear separation towards the green-orange (350-600 nm) for LG, and orange-red (550-725 nm) for PI on the fluorescence spectrum-zone channels. Both dyes showed by non-parametric Kolmcgorov-Smirnov, 99% of probability of difference between heat-killed and live microorganism's histogram-population, with a D = ranking from 0.50-0.64 for LG to 0.91-0.98 for PI. Higher cost and carcinogenic risk when handling PI, make LG a better choice for flow cytometry yeast viability tests.
通过微生物流式细胞术对两种不同染色方法评估酵母活力进行了比较研究。将众所周知的核酸结合活性染料碘化丙啶（PI）与淡绿色（LG）进行比较，淡绿色是一种与细胞质功能相关的活性染料，广泛用于组织学、组织化学和免疫细胞化学染色。所使用的酵母模型是热灭活的季也蒙毕赤酵母细胞。在点图流式细胞仪分析中，染色的热灭活细菌群体在荧光光谱区通道上，对于LG向绿橙色（350 - 600 nm）清晰分离，对于PI向橙红色（550 - 725 nm）清晰分离。两种染料通过非参数Kolmcgorov - Smirnov检验显示，热灭活和活微生物直方图群体之间差异的概率为99%，LG的D值排名从0.50 - 0.64到PI的0.91 - 0.98。处理PI时成本较高且有致癌风险，这使得LG成为流式细胞术酵母活力测试的更好选择。
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