应用激光共聚焦显微技术研究荧光双标记条件下的DAPI光转化效应

Study on the Photoconversion Effect of DAPI under Fluo-rescent Double Labeling Condition by Confocal Microscopy

DAPI作为重要的荧光染料,在细胞核标记实验中被广泛使用。近期有报道指出,DAPI会因紫外光照射而发生光转化效应,光转化后的DAPI可被蓝色光激发而发出绿色荧光信号,进而影响荧光观察。本文通过对细胞核DAPI染色的同时进行细胞质绿色荧光标记,观察到DAPI光转化效率显著下降,说明胞质的绿色荧光对胞核DAPI光转化效应产生抑制。该研究通过激光共聚焦显微手段,对DAPI光转化效应进行观察和分析,为多标记荧光实验观察提供一定依据。

As an important fluorescent dye,DAPI is widely used in nuclear labelingexperiments.Recently,it has been reported that DAPI will produce lightconversion effect due to exposure to UV.After light conversion,DAPI can beexcited by blue light and emit green fluorescence signal,thus affecting thefluorescence experimental observation.In this paper,the nuclear dye DAPI andgreen fluorescence were labeled in the same cells,and it was observed that thelight conversion efficiency of DAPI was significantly reduced,indicating thatthe green fluorescence of cytoplasm had an inhibitory effect on the lightconversion effect of cell nucleus DAPI.In this study,the light conversion effectof DAPI was observed and analyzed through Laser Scanning Confocal Microscope,providing foundation for the multi-labeled fluorescence experimentalobservations that DAPI participated in the same time.