荧光素二乙酸酯荧光微孔板法在细胞活力测定中的应用

Fluorescein Diacetate Microplate Assay in Cell Viability Detection

目的评估荧光素二乙酸酯（FDA）微孔板检测法在细胞活力测定中的应用。方法培养板接种细胞,加入FDA孵育一定时间,多功能微板分析仪测定荧光值。评价FDA孵育时间、浓度及其他影响因素对检测精确度的影响,并与四甲基偶氮唑盐（MTT）方法在H2O2损伤及细胞数量的检测方面进行比较。结果随着孵育时间的延长,荧光值与细胞数的相关系数随之增加,27-30 min时可达到0.99。FDA终浓度在10-30μg/ml时,荧光值与细胞数的相关系数较高。在H2O2损伤模型检测中,与四甲基偶氮唑盐相比,FDA在H2O2高浓度时相关性更强。结论 FDA法检测细胞活力稳定可靠,适用于多种条件下的细胞活力检测及药理学研究。

Objective To investigate the application of the fluorescein diacetate（ FDA） microplate assay in cell viability detection. Methods Cells were seeded in a 96-well culture plate until detection. After incubated with FDA,the plate was detected by fluorescence microplate analyzer. The effects of FDA incubation duration,concentration,and other factors on the assay＇s accuracy and stability were assessed. We also compared the results of FDA with methyl thiazolyl（ MTT） in terms of cell numbers and H2O2 injury. Results Within 0-30 minutes,the fluorescence-cell number coefficient of FDA assay increased with duration and reached 0. 99 in27-30 minutes. The optimum concentration of final FDA in this study was 10-30 μg / ml. On cell viability detection,the result of FDA method was equivalent to MTT method. As to H2O2 injury assay,the sensitivity of FDA method was superior to MTT on the higher concentration H2O2 treatment due to a relative shorter duration for detection. Conclusion As a stable and reliable method,FDA is feasible for cell variability detection under varied conditions.