Influences of Acidic Conditions on Formazan Assay: A Cautionary Note
Influences of Acidic Conditions on Formazan Assay: A Cautionary Note
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DOI：
10.1007/s12010-010-8934-z
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发表时间：
2010-11-01
影响因子：
3
通讯作者：
Kitamura, Masanori
中科院分区：
文献类型：
--
作者：
Johno, Hisashi;Takahashi, Shuhei;Kitamura, Masanori
福尔马赞测定法已用于评价真核和原核细胞的代谢活性几十年。特别是，它经常被应用于在酸性环境下对活细胞进行定量评估，例如，缺血和缺氧。然而，很少有人注意到酸性pH值对甲瓒测定的影响。我们发现，酸性培养条件显着影响的测定结果。四唑-甲的吸光度以pH依赖性方式降低，而不影响细胞活力。这种非特异性效应归因于酸性pH对甲瓒产生的影响。将培养基更换为生理pH的新鲜培养基部分克服了这个问题。在各种实验条件下使用甲瓒测定法评估活细胞时，应仔细考虑酸性培养条件的影响。
Formazan assay has been used for several decades to evaluate metabolic activity of eukaryotic and prokaryotic cells. In particular, it has been often applied for quantitative assessment of viable cells under acidic circumstances caused by, e.g., ischemia and hypoxia. However, little attention has been paid to the influence of acidic pH on formazan assays. We found that acidic culture conditions significantly affect outcomes of the assays. Absorbance of tetrazolium-formazan decreased in a pH-dependent manner without affecting cell viability. This nonspecific effect was ascribed to influences of acidic pH on the production of formazan. Replacement of culture media to fresh medium at physiologic pH partially overcame this problem. The influence of acidic culture conditions should be carefully considered when formazan assays are used for the assessment of viable cells under various experimental situations.