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

DOI: 10.12677/BIPHY.2019.73033, PP. 27-33

Keywords: DAPI，光转化，紫外光，激光共聚焦显微术
DAPI, Photoconversion, Ultraviolet Light, Laser Scanning Confocal Microscope

Full-Text   Cite this paper   Add to My Lib

Abstract:

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 labeling experiments. Recently, it has been reported that DAPI will produce light conversion effect due to exposure to UV. After light conversion, DAPI can be excited by blue light and emit green fluorescence signal, thus affecting the fluorescence experimental observation. In this paper, the nuclear dye DAPI and green fluorescence were labeled in the same cells, and it was observed that the light conversion efficiency of DAPI was significantly reduced, indicating that the green fluorescence of cytoplasm had an inhibitory effect on the light conversion effect of cell nucleus DAPI. In this study, the light conversion effect of DAPI was observed and analyzed through Laser Scanning Confocal Microscope, providing foundation for the multi-labeled fluorescence experimental observations that DAPI participated in the same time.

References

[1]	Kapuscinski, J. (1995) DAPI: A DNA-Specific Fluorescent Probe. Biotechnic & Histochemistry, 70, 220-233. https://doi.org/10.3109/10520299509108199
[2]	Dann, O., Bergen, G., Demant, E. and Volz, G. (1971) Trypanocide Diamidine des 2-Phenyl-benzofurans, 2-Phenyl-indens und 2-Phenyl-indols. Justus Liebigs Annalen der Chemie, 749, 68-89. https://doi.org/10.1002/jlac.19717490110
[3]	Zeman, L.J. and Lusena, C.V. (1975) Preferential Digestion of (A + T)-Rich Stretches of Yeast Mitochondrial DNA in Isolated Mitochondria. European Journal of Biochemistry, 57, 561-567. https://doi.org/10.1111/j.1432-1033.1975.tb02331.x
[4]	Zurek-Biesiada, D., Kedracka-Krok, S. and Dobrucki, J.W. (2013) UV-Activated Conversion of Hoechst 33258, DAPI, and Vybrant DyeCycle Fluorescent Dyes into Blue-Excited, Green-Emitting Protonated Forms. Cytometry Part A, 83A, 441-451. https://doi.org/10.1002/cyto.a.22260
[5]	Karg, T.J. and Golic, K.G. (2018) Photoconversion of DAPI and Hoechst Dyes to Green and Red-Emitting Forms after Exposure to UV Excitation. Chromosoma, 127, 235-245. https://doi.org/10.1007/s00412-017-0654-5
[6]	Piterburg, M., Panet, H. and Weiss, A. (2012) Photoconversion of DAPI Following UV or Violet Excitation Can Cause DAPI to Fluoresce with Blue or Cyan Excitation. Journal of Microscopy, 246, 89-95. https://doi.org/10.1111/j.1365-2818.2011.03591.x
[7]	？urek-Biesiada, D., Waligórski, P. and Dobrucki, J.W. (2014) UV-Induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258. Journal of Fluorescence, 24, 1791-1801. https://doi.org/10.1007/s10895-014-1468-y

Full-Text