Żurek-Biesiada Dominika, Waligórski Piotr, Dobrucki Jurek W
Division of Cell Biophysics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30-387, Kraków, Poland.
J Fluoresc. 2014 Nov;24(6):1791-801. doi: 10.1007/s10895-014-1468-y. Epub 2014 Oct 14.
DNA-bound Hoechst 33258 is readily excited with UV light and emits blue fluorescence, however, upon exposure to UV, the dye undergoes photobleaching as well as photoconversion to a blue-excited green-emitting form. We demonstrate that the UV-generated green-emitting form of Hoechst 33258 exhibits spectral properties very similar to the form of the dye that can be obtained by subjecting it to an acidic environment (pH 0.5-3.0). We also demonstrate that exposure of Hoechst 33258 to UV light (or hydrogen peroxide) leads to generation of the protonated (1+, 2+, 3+ and possibly the 4+) forms of the dye. Photoconversion of Hoechst 33258 has recently been exploited in single molecule localisation microscopy, thus understanding photophysics of this process can facilitate further development of high resolution optical imaging.
与DNA结合的Hoechst 33258很容易被紫外光激发并发出蓝色荧光，然而，在暴露于紫外光时，该染料会发生光漂白以及光转化为蓝色激发绿色发射形式。我们证明，紫外光产生的Hoechst 33258绿色发射形式表现出的光谱特性与通过将其置于酸性环境（pH 0.5 - 3.0）中获得的染料形式非常相似。我们还证明，将Hoechst 33258暴露于紫外光（或过氧化氢）会导致产生该染料的质子化（1 +、2 +、3 +以及可能的4 +）形式。Hoechst 33258的光转化最近已被用于单分子定位显微镜，因此了解这一过程的光物理性质有助于高分辨率光学成像的进一步发展。
