Dorsey J, Yentsch C M, Mayo S, McKenna C
Bigelow Laboratory for Ocean Sciences, West Boothbay Harbor, Maine 04575.
Cytometry. 1989 Sep;10(5):622-8. doi: 10.1002/cyto.990100518.
A standard method for the assessment of cell viability has been developed for marine phytoplankton using an inexpensive stain, fluorescein diacetate (FDA), at .75 microM for 10 min. A flow cytometer was used as the fluorescence detector, providing an assessment of viability for each individual particle. Cell size and chlorophyll fluorescence per cell were assessed simultaneously, permitting an assignment of viability to specific subpopulations, thus increasing the power of the technique. A reasonable correspondence between FDA mean fluorescence intensity per cell and an independent metabolic indicator, photosynthetic capacity measured by 14C, was found. Both FDA mean fluorescence intensity and photosynthetic capacity vary as a function of cell volume. Recovery after extended periods of darkness indicate that cells that are FDA negative may not be dead, but merely quiescent or inactive.
已开发出一种用于评估海洋浮游植物细胞活力的标准方法，该方法使用一种廉价的染料——.75微摩尔的荧光素二乙酸酯（FDA），处理10分钟。使用流式细胞仪作为荧光检测器，可对每个单独的颗粒进行活力评估。同时评估细胞大小和每个细胞的叶绿素荧光，从而能够将活力分配给特定的亚群，进而提高了该技术的效能。发现每个细胞的FDA平均荧光强度与一个独立的代谢指标——通过14C测量的光合能力之间存在合理的对应关系。FDA平均荧光强度和光合能力均随细胞体积而变化。长时间黑暗后的恢复情况表明，FDA呈阴性的细胞可能并未死亡，而仅仅是静止或无活性的。
