两种荧光图像采集设备在采集快速变化荧光图像方面的初步比较

Preliminary Comparative Study of Two Kinds of Devices on Collecting the Quickly Changing Fluorescent Images

目的通过对激光扫描共聚焦显微镜和CCD荧光显微镜采集的DCF探针荧光图像质量的比较,选择适合于快速变化荧光的图像采集设备。方法应用荧光探针H2DCF-DA孵育ECV304细胞后(孵育浓度为10μmol/L,孵育时间30min),分别利用激光扫描共聚焦显微镜和CCD荧光显微镜采集DCF荧光图像,并比较两者的成像质量。结果随着光照致活性氧的产生,DCF荧光的变化速度较快。激光扫描共聚焦显微镜采集的荧光图像表现为细胞内整体的DCF荧光强度增高,图像对比度稍差;而CCD荧光显微镜清晰地呈现细胞内的细节信息,准确地反映了细胞内DCF早期的分布位置。结论在采集快速变化的荧光图像时,CCD荧光显微镜系统优于激光扫描共聚焦显微镜系统。

Objective To choose the suitable device for collecting quickly changing fluorescent image by comparing the quality of the fluorescent image of DCF collected by laser scanning confocal microscope and fluorescent microscope with charge-coupled device (CCD). Methods ECV 304 cells were subcultured for 24 h. H2DCF-DA was incubated for 30 min with the final concentration of 10μmol/L. DCF was generated during the procedure of light irradiation. The fluorescent images of DCF were collected spontaneously by laser scanning confocal microscope and CCD fluorescent microscope respectively. Results The fluorescence intensity of DCF changed rapidly with the generation of reactive oxygen species induced by light irradiation. The fluorescent images of DCF collected by CCD fluorescent microscope was clearer than that by laser scanning confocal microscope. Conclusions CCD fluorescent microscope is superior to laser scanning confocal microscope in collecting the quickly changing fluores- cent image such as DCF.