Autofluorescence of chloroplasts measured by a laser scanning confocal microscope
FENG Qianqian1,2, WANG Wenjuan1,3, LI Haidong1,2, PAN Xun1,2
1. School of Life Sciences, Tsinghua University, Beijing 100084, China;
2. Center of Biomedical Analysis, Tsinghua University, Beijing 100084, China;
3. Technology Center for Protein Sciences, Tsinghua University, Beijing 100084, China
Abstract：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 autofluorescence 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.
冯倩倩, 王文娟, 李海冬, 潘勋. 利用激光扫描共聚焦显微镜研究叶绿体自发荧光[J]. 清华大学学报（自然科学版）, 10.16511/j.cnki.qhdxxb.2017.26.034.
FENG Qianqian, WANG Wenjuan, LI Haidong, PAN Xun. Autofluorescence of chloroplasts measured by a laser scanning confocal microscope. Journal of Tsinghua University(Science and Technology), 2017, 57(6): 651-654,660.
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