Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity
Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity
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DOI：
10.1046/j.1432-1327.2000.01606.x
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发表时间：
2000-09-01
期刊：
影响因子：
--
通讯作者：
Pognan, F
中科院分区：
文献类型：
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作者：
O'Brien, J;Wilson, I;Pognan, F
我们在这里展示了Alfred Blue作为刃天青的身份。“刃天青还原试验”已被用于监测牛奶中的细菌和酵母污染，以及评估精液质量约50年。刃天青（蓝色和非荧光）被还原为试卤灵（粉红色和高荧光），其进一步还原为氢化试卤灵（无色和非荧光）。目前尚不清楚这种还原是如何发生的，细胞内通过酶活性或在培养基中作为化学反应，尽管在细胞质和死细胞的活细胞核中发现了还原荧光形式的Alcohol Blue。最近，染料作为测量细胞增殖和细胞毒性的非常简单和通用的方法而受到欢迎。这种染料与其他细胞毒性或增殖试验相比具有许多优点，但我们观察到Alcohol Blue常规使用的几个缺点。在不同细胞系和大鼠原代肝细胞中使用几种毒物进行的试验表明，培养基中Alcohol Blue的荧光产物蓄积，这可能导致高估细胞群。此外，代谢活跃的细胞大量减少Alcohol Blue导致最终的非荧光产物，因此低估了细胞活性。
We show here the identity of Alamar Blue as resazurin. The 'resazurin reduction test' has been used for about 50 years to monitor bacterial and yeast contamination of milk, and also for assessing semen quality. Resazurin (blue and nonfluorescent) is reduced to resorufin (pink and highly fluorescent) which is further reduced to hydroresorufin (uncoloured and nonfluorescent). It is still not known how this reduction occurs, intracellularly via enzyme activity or in the medium as a chemical reaction, although the reduced fluorescent form of Alamar Blue was found in the cytoplasm and of living cells nucleus of dead cells. Recently, the dye has gained popularity as a very simple and versatile way of measuring cell proliferation and cytotoxicity. This dye presents numerous advantages over other cytotoxicity or proliferation tests but we observed several drawbacks to the routine use of Alamar Blue. Tests with several toxicants in different cell lines and rat primary hepatocytes have shown accumulation of the fluorescent product of Alamar Blue in the medium which could lead to an overestimation of cell population. Also, the extensive reduction of Alamar Blue by metabolically active cells led to a final nonfluorescent product, and hence an underestimation of cellular activity.