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Chemically Induced Photoswitching of Fluorescent Probes—A General Concept for Super-Resolution Microscopy
Biotechnology & Biophysics, Julius-Maximilians-University Würzburg, Am Hubland, 97074 Würzburg, Germany
Bioquant Centre, University of Heidelberg, INF 297, 69120 Heidelberg, Germany
Department of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany
* Author to whom correspondence should be addressed.
Received: 18 March 2011; in revised form: 8 April 2011 / Accepted: 12 April 2011 / Published: 13 April 2011
Abstract: We review fluorescent probes that can be photoswitched or photoactivated and are suited for single-molecule localization based super-resolution microscopy. We exploit the underlying photochemical mechanisms that allow photoswitching of many synthetic organic fluorophores in the presence of reducing agents, and study the impact of these on the photoswitching properties of various photoactivatable or photoconvertible fluorescent proteins. We have identified mEos2 as a fluorescent protein that exhibits reversible photoswitching under various imaging buffer conditions and present strategies to characterize reversible photoswitching. Finally, we discuss opportunities to combine fluorescent proteins with organic fluorophores for dual-color photoswitching microscopy.
Keywords: photoswitchable organic fluorophores; fluorescent proteins; super-resolution; PALM; dSTORM
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Endesfelder, U.; Malkusch, S.; Flottmann, B.; Mondry, J.; Liguzinski, P.; Verveer, P.J.; Heilemann, M. Chemically Induced Photoswitching of Fluorescent Probes—A General Concept for Super-Resolution Microscopy. Molecules 2011, 16, 3106-3118.
Endesfelder U, Malkusch S, Flottmann B, Mondry J, Liguzinski P, Verveer PJ, Heilemann M. Chemically Induced Photoswitching of Fluorescent Probes—A General Concept for Super-Resolution Microscopy. Molecules. 2011; 16(4):3106-3118.
Endesfelder, Ulrike; Malkusch, Sebastian; Flottmann, Benjamin; Mondry, Justine; Liguzinski, Piotr; Verveer, Peter J.; Heilemann, Mike. 2011. "Chemically Induced Photoswitching of Fluorescent Probes—A General Concept for Super-Resolution Microscopy." Molecules 16, no. 4: 3106-3118.