Department of Biotechnology and Biophysics, Biocenter, Julius Maximilian University of Würzburg, Würzburg, Germany.
Chemical Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD, USA.
Nat Methods. 2021 Mar;18(3):253-257. doi: 10.1038/s41592-021-01061-2. Epub 2021 Feb 25.
Illumination of fluorophores can induce a loss of the ability to fluoresce, known as photobleaching. Interestingly, some fluorophores photoconvert to a blue-shifted fluorescent molecule as an intermediate on the photobleaching pathway, which can complicate multicolor fluorescence imaging, especially under the intense laser irradiation used in super-resolution fluorescence imaging. Here, we discuss the mechanisms of photoblueing of fluorophores and its impact on fluorescence imaging, and show how it can be prevented.
荧光团的激发会导致其丧失荧光能力,这被称为光漂白。有趣的是,一些荧光团在光漂白途径中会光转化为一个蓝移的荧光分子作为中间产物,这可能会使多色荧光成像复杂化,特别是在超分辨率荧光成像中使用的高强度激光照射下。在这里,我们讨论了荧光团光漂白的机制及其对荧光成像的影响,并展示了如何预防它。