Gray D W, Morris P J
Nuffield Department of Surgery, University of Oxford, John Radcliffe Hospital, Headington, England.
Stain Technol. 1987 Nov;62(6):373-81. doi: 10.3109/10520298709108028.
There is a need for a simple, rapid, sensitive method for assessing the viability of isolated islets of Langerhans. In this study the fluorescent dyes fluorescein diacetate (FDA) and ethidium bromide (EB) have been used to provide a viability assay for isolated rat islets. Discrimination of living from dead islets is efficient; in a blind sorting experiment using freshly isolated islets and islets killed by either heat or alcohol, viability determined by insulin secretion in response to glucose stimulation correlated well with viability as determined by FDA/EB staining. Furthermore, it is possible to discriminate degrees of viability, and a scoring system is described for this purpose which is shown to correlate with another index of viability, the ATP content. A reliable viability stain should not itself be toxic; FDA/EB stained islets remain viable after staining, showing normal response to glucose stimulation and normal function after transplantation.
需要一种简单、快速、灵敏的方法来评估分离的胰岛的活力。在本研究中，荧光染料荧光素二乙酸酯（FDA）和溴化乙锭（EB）已被用于为分离的大鼠胰岛提供活力测定。区分活胰岛和死胰岛的效果良好；在一项使用新鲜分离的胰岛以及经加热或酒精处理致死的胰岛进行的盲选实验中，通过对葡萄糖刺激的胰岛素分泌所测定的活力与通过FDA/EB染色所测定的活力高度相关。此外，有可能区分活力程度，并为此描述了一种评分系统，该系统显示与另一个活力指标ATP含量相关。一种可靠的活力染色剂本身不应具有毒性；经FDA/EB染色的胰岛在染色后仍保持活力，对葡萄糖刺激表现出正常反应，移植后功能也正常。
