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Transient state imaging of live cells using single plane illumination and arbitrary duty cycle excitation pulse trains
Author(s) -
Mücksch Jonas,
Spielmann Thiemo,
Sisamakis Evangelos,
Widengren Jerker
Publication year - 2015
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201400015
Subject(s) - duty cycle , transient (computer programming) , excitation , pulse (music) , fluorophore , optics , physics , frame rate , plane (geometry) , biological system , fluorescence , computer science , detector , mathematics , geometry , quantum mechanics , biology , operating system , power (physics)
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)