Major signal increase in fluorescence microscopy through dark-state relaxation.
We report a substantial signal gain in fluorescence microscopy by ensuring that transient molecular dark states with lifetimes >1 micros, 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 fluor...
| Main Authors: | , , |
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| פורמט: | Journal article |
| שפה: | English |
| יצא לאור: |
2007
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| _version_ | 1826265319472103424 |
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| author | Donnert, G Eggeling, C Hell, S |
| author_facet | Donnert, G Eggeling, C Hell, S |
| author_sort | Donnert, G |
| collection | OXFORD |
| description | We report a substantial signal gain in fluorescence microscopy by ensuring that transient molecular dark states with lifetimes >1 micros, 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 high-repetition-rate pulsed illumination was replaced with pulses featuring temporal pulse separation >1 micros. 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. |
| first_indexed | 2024-03-06T20:21:49Z |
| format | Journal article |
| id | oxford-uuid:2e0ea7cb-1a18-4fe5-81a7-811b0aae0d30 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T20:21:49Z |
| publishDate | 2007 |
| record_format | dspace |
| spelling | oxford-uuid:2e0ea7cb-1a18-4fe5-81a7-811b0aae0d302022-03-26T12:46:45ZMajor signal increase in fluorescence microscopy through dark-state relaxation.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:2e0ea7cb-1a18-4fe5-81a7-811b0aae0d30EnglishSymplectic Elements at Oxford2007Donnert, GEggeling, CHell, SWe report a substantial signal gain in fluorescence microscopy by ensuring that transient molecular dark states with lifetimes >1 micros, 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 high-repetition-rate pulsed illumination was replaced with pulses featuring temporal pulse separation >1 micros. 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. |
| spellingShingle | Donnert, G Eggeling, C Hell, S Major signal increase in fluorescence microscopy through dark-state relaxation. |
| title | Major signal increase in fluorescence microscopy through dark-state relaxation. |
| title_full | Major signal increase in fluorescence microscopy through dark-state relaxation. |
| title_fullStr | Major signal increase in fluorescence microscopy through dark-state relaxation. |
| title_full_unstemmed | Major signal increase in fluorescence microscopy through dark-state relaxation. |
| title_short | Major signal increase in fluorescence microscopy through dark-state relaxation. |
| title_sort | major signal increase in fluorescence microscopy through dark state relaxation |
| work_keys_str_mv | AT donnertg majorsignalincreaseinfluorescencemicroscopythroughdarkstaterelaxation AT eggelingc majorsignalincreaseinfluorescencemicroscopythroughdarkstaterelaxation AT hells majorsignalincreaseinfluorescencemicroscopythroughdarkstaterelaxation |