RELIABILITY OF CALCEIN ACETOXY METHYL-ESTER AND ETHIDIUM HOMODIMER OR PROPIDIUM IODIDE FOR VIABILITY ASSESSMENT OF MICROBES

RELIABILITY OF CALCEIN ACETOXY METHYL-ESTER AND ETHIDIUM HOMODIMER OR PROPIDIUM IODIDE FOR VIABILITY ASSESSMENT OF MICROBES
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DOI:
10.1016/s0167-7012(93)80010-4
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发表时间:
1993-01-01
影响因子:
2.2
通讯作者:
CUSHION, MT
CUSHION, MT
中科院分区:
生物学4区
文献类型:
--
作者:
KANESHIRO, ES;WYDER, MA;CUSHION, MT

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酯酶指示剂,钙黄绿素乙酰氧基甲酯,与乙锭同型二聚体或碘化丙啶组合使用,以评估其适用性的真核和原核微生物的活力测定。这些荧光探针先前被证明可用于确定培养物中哺乳动物细胞的活力,但未评估其在测试各种微生物中的可靠性。荧光强度、缺乏漂白和对pH差异不敏感是钙黄绿素乙酰氧基甲酯的特征,这使得能够在足够高的放大倍数下进行常规彩色显微摄影记录,以观察细胞形态,从而正确鉴定微生物。该方法应用于原生动物,染色结果与存活率具有良好的相关性。相比之下,该方法对于测试的酵母和细菌细胞的评估不可靠。
An esterase indicator, calcein acetoxy methyl ester, was used in combination with ethidium homodimer or propidium iodide to evaluate its applicability for viability assays of eukaryotic and prokaryotic microorganisms. These fluorescent probes were previously proven useful in determining the viability of mammalian cells in culture but were not evaluated for their reliability in testing various microorganisms. The fluorescence intensity, lack of bleaching and insensitivity to differences in pH that are characteristic of calcein acetoxy methyl ester enable routine color photomicrographic documentation at magnifications high enough to observe cell morphology for proper identification of the microorganisms. There was good correlation of staining and viability when the method was applied to protozoans. In contrast, the method was not reliable for assessment of the yeast and bacterial cells tested.