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Structure and mechanism of the reversible photoswitch of a fluorescent protein
Proteins that can be reversibly photoswitched between a fluorescent and a nonfluorescent state bear enormous potential in diverse fields, such as data storage, in vivo protein tracking, and subdiffraction...
PublicationsBreaking the diffraction barrier in fluorescence microscopy at low light intensities by using reversibly photoswitchable proteins
Fluorescence microscopy is indispensable in many areas of science, but until recently, diffraction has limited the resolution of its lens-based variant. The diffraction barrier has been broken by a saturated...
PublicationsMacromolecular-scale resolution in biological fluorescence microscopy
We demonstrate far-field fluorescence microscopy with a focal-plane resolution of 15-20 nm in biological samples. The 10- to 12-fold multilateral increase in resolution below the diffraction barrier has...
PublicationsStructural basis for reversible photoswitching in Dronpa
Dronpa is a novel GFP-like fluorescent protein with exceptional light-controlled switching properties. It may be reversibly switched between a fluorescent on-state and a nonfluorescent off-state by irradiation...
PublicationsFast molecular tracking maps nanoscale dynamics of plasma membrane lipids
We describe an optical method capable of tracking a single fluorescent molecule with a flexible choice of high spatial accuracy (similar to 10-20 nm standard deviation or similar to 20-40 nm full-width-at-half-maximum)...
PublicationsSTED Nanoscopy with Time-Gated Detection: Theoretical and Experimental Aspects
In a stimulated emission depletion (STED) microscope the region in which fluorescence markers can emit spontaneously shrinks with continued STED beam action after a singular excitation event. This fact...
PublicationsPeroxisomal Import Reduces the Proapoptotic Activity of Deubiquitinating Enzyme USP2
The human deubiquitinating enzyme ubiquitin-specific protease 2 (USP2) regulates multiple cellular pathways, including cell proliferation and apoptosis. As a result of alternative splicing four USP2 isoenzymes...
PublicationsFluorescence fluctuation spectroscopy in subdiffraction focal volumes
We establish fluorescence fluctuation spectroscopy (FFS) with nanoscale detection volumes generated by stimulated emission depletion. Our method applies fluorescence correlation spectroscopy and fluorescence...
PublicationsBreaking the diffraction barrier in fluorescence microscopy by optical shelving
We report the breaking of the diffraction resolution barrier in far-field fluorescence microscopy by transiently shelving the fluorophore in a metastable dark state. Using a relatively modest light intensity...
PublicationsTriplet-relaxation microscopy with bunched pulsed excitation
Obtaining high signal levels in fluorescence microscopy is usually spoiled by the concomitant population of the dark ( triplet) state of the marker, which is often followed by photobleaching. Recently,...
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