Major signal increase in fluorescence microscopy through dark-state relaxation
2007 | journal article; research paper. A publication with affiliation to the University of Göttingen.
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- Authors
- Donnert, Gerald; Eggeling, Christian ; Hell, Stefan
- Abstract
- 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.
- Issue Date
- 2007
- Journal
- Nature Methods
- ISSN
- 1548-7091
- Language
- English