Krause A W, Carley W W, Webb W W
J Histochem Cytochem. 1984 Oct;32(10):1084-90. doi: 10.1177/32.10.6090533.
Erythrosin B and trypan blue are tested and compared for their effectiveness as vital exclusion stains for mammalian cells in monolayer culture. Both stains are supposed to mark cells that have lost membrane integrity. Fluorescein diacetate (FDA), an efficient vital inclusion stain, is used as a control, as it marks cells retaining membrane integrity. Erythrosin B and FDA are used as fluorescent dyes, whereas trypan blue colors via light absorption. The effectiveness of both vital exclusion stains is assayed by their ability to stain a high percentage of monolayer cells exposed to treatments lethal to an entire cell population. Two types of lethal treatment, severe heat and metabolic poison, are employed. Erythrosin B stains all monolayer cells immediately after complete lethal treatment. Trypan blue optimally stains only about 60% of monolayer cells. Cell staining by erythrosin B and by FDA are found to be mutually exclusive. This result demonstrates the coincidence of viability indications by erythrosin B and FDA and thus confirms the reliability of both viability stains as they probe membrane permeability via independent mechanisms. This study shows that erythrosin B is an effective, nontoxic, and convenient fluorescent vital exclusion dye for three mammalian cell lines in monolayer culture, but tends to disqualify trypan blue for this application.
对赤藓红B和台盼蓝作为哺乳动物单层培养细胞的活性排斥染料的有效性进行了测试和比较。两种染料都应标记失去膜完整性的细胞。荧光素二乙酸酯（FDA）是一种有效的活性摄入染料，用作对照，因为它标记保留膜完整性的细胞。赤藓红B和FDA用作荧光染料，而台盼蓝通过光吸收显色。两种活性排斥染料的有效性通过它们对暴露于对整个细胞群体致死的处理的单层细胞进行高比例染色的能力来测定。采用了两种致死处理方式，即高温和代谢毒物。在完全致死处理后，赤藓红B立即对所有单层细胞进行染色。台盼蓝最佳情况下仅能对约60%的单层细胞进行染色。发现赤藓红B和FDA对细胞的染色相互排斥。这一结果证明了赤藓红B和FDA的活力指示结果一致，从而证实了这两种活力染料在通过独立机制探测膜通透性方面的可靠性。本研究表明，赤藓红B是一种有效、无毒且方便的荧光活性排斥染料，适用于三种哺乳动物单层培养细胞系，但台盼蓝在该应用中可能不适用。
