利用激光扫描共聚焦显微镜研究叶绿体自发荧光

Autofluorescence of chloroplasts measured by a laser scanning confocal microscope

该文利用激光扫描共聚焦显微镜研究烟草叶片叶绿体自发荧光,为叶绿体研究提供理论和实验依据。首先,利用不同波长激发光的光谱扫描功能分别激发烟草叶片叶绿体,得到叶绿体自发荧光光谱;其次,利用不同激发光激发烟草叶片叶绿体,获取叶绿体自发荧光图像,并对荧光强度进行定量分析比较;最后,利用488nm激发光漂白烟草叶片叶绿体,观察自发荧光的稳定性。实验结果显示:488nm激光对叶绿体的激发效率最高,561nm激光的激发效率最低;发射光谱集中在大于637nm的波段,峰值在681nm处;且叶绿体自发荧光非常稳定。激光扫描共聚焦显微镜能够准确研究叶绿体自发荧光,对研究植物组织成像提供重要帮助,也对叶绿体基因工程研究有深远影响。

The autofluorescence of nicotiana benthamiana chloroplasts was studied using a laser scanning confocal microscope. The chloroplast autofluorescence was obtained by spectral scanning with various excitation wavelengths. Then, chloroplast autofluorescence images were obtained using laser scanning confocal microscopy with various excitation wavelengths with intensity quantification. Finally, nicotiana benthamiana chloroplasts were illuminated with 488 nm light to test the autofluorescence stability. 488 nm light has the highest excitation efficiency for chloroplast autofluorescence, while 561 nm light has the lowest excitation efficiency. The autofluorescence chloroplast emission spectrum starts from 637 nm and peaks at 681 nm. The measurements show that the chloroplast autoiluorescence is very stable and that laser scanning confocal microscopy can accurately measure chloroplasts autofluorescence. This work will facilitate improved plant tissue imaging chloroplast genetic engineering research.