Laboratory of Experimental Biophysics, Institute of Physics of Biological Systems, École Polytechnique Fédérale de Lausanne (EPFL), National Centre of Competence in Research (NCCR) in Chemical Biology, Lausanne (Switzerland).
Chemphyschem. 2014 Mar 17;15(4):750-5. doi: 10.1002/cphc.201301004. Epub 2014 Feb 19.
Cell-permeable rhodamine dyes are reductively quenched by NaBH4 into a non-fluorescent leuco-rhodamine form. Quenching is reversible, and their fluorescence is recovered when the dyes are oxidized. In living cells, oxidation occurs spontaneously, and can result in up to ten-fold higher densities of single molecule localizations, and more photons per localization as compared with unmodified dyes. These two parameters directly impact the achievable resolution, and we see a significant improvement in the quality of live-cell point-localization super-resolution images taken with reduced dyes. These improvements carry over to increase the density of trajectories for single-molecule tracking experiments.
具有细胞通透性的罗丹明染料可被 NaBH4 还原猝灭为非荧光的隐色罗丹明形式。猝灭是可逆的，当染料被氧化时，其荧光会恢复。在活细胞中，氧化会自发发生，与未修饰的染料相比，可导致单分子定位的密度增加十倍以上，并且每个定位的光子数增加。这两个参数直接影响可实现的分辨率，我们观察到使用还原染料拍摄的活细胞点定位超分辨率图像的质量有了显著提高。这些改进可提高单分子跟踪实验轨迹的密度。
