Transient state imaging of live cells using single plane illumination and arbitrary duty cycle excitation pulse trains |
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| Authors: | Jonas Mücksch Thiemo Spielmann Evangelos Sisamakis Jerker Widengren |
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| Affiliation: | Experimental Biomolecular Physics, Department of Applied Physics, Royal Institute of Technology, Albanova University Center, Stockholm, Sweden |
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| Abstract: | We demonstrate the applicability of Single Plane Illumination Microscopy to Transient State Imaging (TRAST), offering sensitive microenvironmental information together with optical sectioning and reduced overall excitation light exposure of the specimen. The concept is verified by showing that transition rates can be determined accurately for free dye in solution and that fluorophore transition rates can be resolved pixel‐wise in live cells. Furthermore, we derive a new theoretical framework for analyzing TRAST data acquired with arbitrary duty cycle pulse trains. By this analysis it is possible to reduce the overall measurement time and thereby enhance the frame rates in TRAST imaging. (© 2014 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim) |
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| Keywords: | fluorescence single plane illumination microscopy transient state imaging triplet state redox state live cell imaging |
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