Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy.

Fluorescent proteins that can be reversibly photoswitched between a fluorescent and a nonfluorescent state are important for innovative microscopy schemes, such as protein tracking, fluorescence resonance energy transfer imaging, sub-diffraction resolution microscopy and others. However, all availab...

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المؤلفون الرئيسيون: Andresen, M, Stiel, A, Fölling, J, Wenzel, D, Schönle, A, Egner, A, Eggeling, C, Hell, S, Jakobs, S
التنسيق: Journal article
اللغة:English
منشور في: 2008
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author Andresen, M
Stiel, A
Fölling, J
Wenzel, D
Schönle, A
Egner, A
Eggeling, C
Hell, S
Jakobs, S
author_facet Andresen, M
Stiel, A
Fölling, J
Wenzel, D
Schönle, A
Egner, A
Eggeling, C
Hell, S
Jakobs, S
author_sort Andresen, M
collection OXFORD
description Fluorescent proteins that can be reversibly photoswitched between a fluorescent and a nonfluorescent state are important for innovative microscopy schemes, such as protein tracking, fluorescence resonance energy transfer imaging, sub-diffraction resolution microscopy and others. However, all available monomeric reversibly switchable fluorescent proteins (RSFPs) have similar properties and switching characteristics, thereby limiting their use. Here, we introduce two bright green fluorescent RSFPs, bsDronpa and Padron, generated by extensive mutagenesis of the RSFP Dronpa, with unique absorption and switching characteristics. Whereas bsDronpa features a broad absorption spectrum extending into the UV, Padron displays a switching behavior that is reversed to that of all green fluorescent RSFPs known to date. These two RSFPs enable live-cell fluorescence microscopy with multiple labels using a single detection color, because they can be distinguished by photoswitching. Furthermore, we demonstrate dual-color fluorescence microscopy with sub-diffraction resolution using bsDronpa and Dronpa whose emission maxima are separated by <20 nm.
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spelling oxford-uuid:4fc3a90a-023c-4a15-937b-89ea19b81d742022-03-26T16:09:25ZPhotoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:4fc3a90a-023c-4a15-937b-89ea19b81d74EnglishSymplectic Elements at Oxford2008Andresen, MStiel, AFölling, JWenzel, DSchönle, AEgner, AEggeling, CHell, SJakobs, SFluorescent proteins that can be reversibly photoswitched between a fluorescent and a nonfluorescent state are important for innovative microscopy schemes, such as protein tracking, fluorescence resonance energy transfer imaging, sub-diffraction resolution microscopy and others. However, all available monomeric reversibly switchable fluorescent proteins (RSFPs) have similar properties and switching characteristics, thereby limiting their use. Here, we introduce two bright green fluorescent RSFPs, bsDronpa and Padron, generated by extensive mutagenesis of the RSFP Dronpa, with unique absorption and switching characteristics. Whereas bsDronpa features a broad absorption spectrum extending into the UV, Padron displays a switching behavior that is reversed to that of all green fluorescent RSFPs known to date. These two RSFPs enable live-cell fluorescence microscopy with multiple labels using a single detection color, because they can be distinguished by photoswitching. Furthermore, we demonstrate dual-color fluorescence microscopy with sub-diffraction resolution using bsDronpa and Dronpa whose emission maxima are separated by <20 nm.
spellingShingle Andresen, M
Stiel, A
Fölling, J
Wenzel, D
Schönle, A
Egner, A
Eggeling, C
Hell, S
Jakobs, S
Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy.
title Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy.
title_full Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy.
title_fullStr Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy.
title_full_unstemmed Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy.
title_short Photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy.
title_sort photoswitchable fluorescent proteins enable monochromatic multilabel imaging and dual color fluorescence nanoscopy
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AT stiela photoswitchablefluorescentproteinsenablemonochromaticmultilabelimaginganddualcolorfluorescencenanoscopy
AT follingj photoswitchablefluorescentproteinsenablemonochromaticmultilabelimaginganddualcolorfluorescencenanoscopy
AT wenzeld photoswitchablefluorescentproteinsenablemonochromaticmultilabelimaginganddualcolorfluorescencenanoscopy
AT schonlea photoswitchablefluorescentproteinsenablemonochromaticmultilabelimaginganddualcolorfluorescencenanoscopy
AT egnera photoswitchablefluorescentproteinsenablemonochromaticmultilabelimaginganddualcolorfluorescencenanoscopy
AT eggelingc photoswitchablefluorescentproteinsenablemonochromaticmultilabelimaginganddualcolorfluorescencenanoscopy
AT hells photoswitchablefluorescentproteinsenablemonochromaticmultilabelimaginganddualcolorfluorescencenanoscopy
AT jakobss photoswitchablefluorescentproteinsenablemonochromaticmultilabelimaginganddualcolorfluorescencenanoscopy