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Int. J. Mol. Sci. 2017, 18(7), 1524; doi:10.3390/ijms18071524

Combining Primed Photoconversion and UV-Photoactivation for Aberration-Free, Live-Cell Compliant Multi-Color Single-Molecule Localization Microscopy Imaging

Department of Systems and Synthetic Microbiology, Max Planck Institute for Terrestrial Microbiology & LOEWE Center for Synthetic Microbiology (SYNMIKRO), Karl-von-Frisch-Str. 16, 35043 Marburg, Germany
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Received: 11 June 2017 / Revised: 4 July 2017 / Accepted: 5 July 2017 / Published: 14 July 2017
(This article belongs to the Special Issue Fluorescent Proteins)
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Abstract

Super-resolution fluorescence microscopy plays a major role in revealing the organization and dynamics of living cells. Nevertheless, single-molecule localization microscopy imaging of multiple targets is still limited by the availability of suitable fluorophore combinations. Here, we introduce a novel imaging strategy which combines primed photoconversion (PC) and UV-photoactivation for imaging different molecular species tagged by suitable fluorescent protein combinations. In this approach, the fluorescent proteins can be specifically photoactivated/-converted by different light wavelengths using PC and UV-activation modes but emit fluorescence in the same spectral emission channel. We demonstrate that this aberration-free, live-cell compatible imaging method can be applied to various targets in bacteria, yeast and mammalian cells and can be advantageously combined with correlative imaging schemes. View Full-Text
Keywords: multi-color imaging; primed conversion; live cell imaging; single-molecule localization microscopy multi-color imaging; primed conversion; live cell imaging; single-molecule localization microscopy
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Virant, D.; Turkowyd, B.; Balinovic, A.; Endesfelder, U. Combining Primed Photoconversion and UV-Photoactivation for Aberration-Free, Live-Cell Compliant Multi-Color Single-Molecule Localization Microscopy Imaging. Int. J. Mol. Sci. 2017, 18, 1524.

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