Autofluorescence of chloroplasts measured by a laser scanning confocal microscope

被引:0
|
作者
Feng Q. [1 ,2 ]
Wang W. [1 ,3 ]
Li H. [1 ,2 ]
Pan X. [1 ,2 ]
机构
[1] School of Life Sciences, Tsinghua University, Beijing
[2] Center of Biomedical Analysis, Tsinghua University, Beijing
[3] Technology Center for Protein Sciences, Tsinghua University, Beijing
来源
Pan, Xun (panx@biomed.tsinghua.edu.cn) | 1600年 / Tsinghua University卷 / 57期
关键词
Autofluorescence; Chloroplast; Laser scanning confocal microscopy; Spectral imaging;
D O I
10.16511/j.cnki.qhdxxb.2017.26.034
中图分类号
学科分类号
摘要
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. © 2017, Tsinghua University Press. All right reserved.
引用
收藏
页码:651 / 654and660
相关论文
共 20 条
  • [1] White Amp J.G., Amos W.B., Confocal microscopy comes of age, Nature, 328, 6126, pp. 183-184, (1987)
  • [2] White J.G., Amos W.B., Fordham M., An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy, Journal of Cell Biology, 105, 1, pp. 41-48, (1987)
  • [3] Turillazzi E., Karch S.B., Neri M., Et al., Confocal laser scanning microscopy: Using new technology to answer old questions in forensic investigations, Deutsche Zeitschrift Für Die Gesamte Gerichtliche Medizin, 122, 2, pp. 173-177, (2008)
  • [4] Wade M.H., Fluorescence quantifi cati on in living cells, Biomed Tech, 87, pp. 43-48, (1992)
  • [5] Zhou T., Yang Y., Zhang D., Et al., Laser scanning confocal microscope and its applications in biomedicine, Bulletin of The Academy of Military Medical Sciences, 1, pp. 69-73, (2002)
  • [6] Hong R., Observation of rice embryo sac development with confocal laser scanning microscopy, Acta Botanica Sinica, 40, 9, pp. 786-789, (1998)
  • [7] Wang D., Wang X., Zhang W., Revealing the F-actin networks in interphase nuclei of garlic clove cells by confocal fluorescence microscopy, Acta Botanica Sinica, 42, 11, pp. 1167-1171, (2000)
  • [8] Lamprecht A., Schafer U.F., Lehr C.M., Characterization of microcapsules by confocal laser scanning microscopy: Structure, capsule wall composition and encapsulation rate, European Journal of Pharmaceutics & Biopharmaceutics, 49, 1, pp. 1-9, (2000)
  • [9] Kubinova L., Janacek J., Karen P., Et al., Confocal stereology and image analysis: Methods for estimating geometrical characteristics of cells and tissues from three-dimensional confocal images, Physiological Research, 53, pp. 47-55, (2004)
  • [10] Qiu L., Li J., Zhao W., Et al., Laser confocal measurement system for curvature radii of lenses, Optics and Precision Engineering, 21, 2, pp. 246-252, (2013)