Major signal increase in fluorescence microscopy through dark-state relaxation
Major signal increase in fluorescence microscopy through dark-state relaxation
复制标题
DOI：
10.1038/nmeth986
复制
发表时间：
2007-01-01
期刊：
影响因子：
48
通讯作者：
Hell, Stefan W.
中科院分区：
文献类型：
--
作者：
Donnert, Gerald;Eggeling, Christian;Hell, Stefan W.
我们报告了大量的信号增益荧光显微镜，确保瞬态分子暗态与寿命> 1 μ s，如三重态放松之间的两个分子吸收事件。对于GFP和罗丹明染料Atto 532，当强连续波或高重复率脉冲照明被具有时间脉冲间隔> 1 μ s的脉冲取代时，我们观察到在分子漂白之前总荧光产率增加5-25倍。观察到单光子和双光子激发的信号增益。在照明过程中服从暗态或三重态弛豫意味着向具有低光漂白和强荧光通量的成像迈出了重要一步。(c)2007年，自然出版集团。
We report a substantial signal gain in fluorescence microscopy by ensuring that transient molecular dark states with lifetimes > 1 mu s, such as the triplet state relax between two molecular absorption events. For GFP and Rhodamine dye Atto532, we observed a 5-25-fold increase in total fluorescence yield before molecular bleaching when strong continuous-wave or highrepetitionratepulsed illumination was replaced with pulses featuring temporal pulse separation > 1 mu s. The signal gain was observed both for one- and two-photon excitation. Obeying dark or triplet state relaxation in the illumination process signifies a major step toward imaging with low photobleaching and strong fluorescence fluxes. (c) 2007 Nature Publishing Group.