Stefan W. Hell
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View article: The near-infrared bacteriophytochrome-derived fluorescent protein PENELOPE enables RESOLFT superresolution microscopy
The near-infrared bacteriophytochrome-derived fluorescent protein PENELOPE enables RESOLFT superresolution microscopy Open
REversible Saturable Optical Linear Fluorescence Transitions (RESOLFT) superresolution microscopy fundamentally overcomes the diffraction barrier in far-field fluorescence microscopy. It relies on reversibly switchable fluorescent proteins…
On‐Target Photoassembly of Pyronin Dyes for Super‐Resolution Microscopy Open
Controlled photoactivation is an auspicious and emerging approach in super‐resolution microscopy, offering virtually zero background signal from the marker prior to activation. Pyronins are well‐established fluorophores, but due to their i…
On‐Target Photoassembly of Pyronin Dyes for Super‐Resolution Microscopy Open
Controlled photoactivation is an auspicious and emerging approach in super‐resolution microscopy, offering virtually zero background signal from the marker prior to activation. Pyronins are well‐established fluorophores, but due to their i…
Supramolecular Complexation of Quenched Rosamines with Cucurbit[7]Uril: Fluorescence Turn-ON Effect for Super-Resolution Imaging Open
Here, we present a fluorogenic supramolecular host-guest system comprising cucurbit[7]uril (CB7) and a rationally designed rosamine fluorescent dye (P-ARose) tailored for super-resolution imaging applications. By strategically designing th…
On-target photoassembly of pyronin dyes for super-resolution microscopy Open
Controlled photoactivation is an auspicious and emerging approach in super-resolution microscopy, offering virtually zero background signal from the marker prior to activation. Pyronins are well-established fluorophores, but due to their i…
Caging-group-free photoactivatable fluorophores with far-red emission Open
State of the art super-resolution microscopy methods profit greatly when combined with photoactivatable or photoswitchable fluorophores with far-red emission, but the majority of these fluorophores require specialized buffers or rely on ph…
Diffraction minima resolve point scatterers at few hundredths of the wavelength Open
Resolving two or more constantly scattering identical point sources using freely propagating waves is limited by diffraction. Here we show that, by illuminating with a diffraction minimum, a given number of point scatterers can be resolved…
Asymmetric Donor–Acceptor 2,7-Disubstituted Fluorenes and Their 9-Diazoderivatives: Synthesis, Optical Spectra and Photolysis Open
In a search for dyes photoactivatable with visible light, fluorenes with substituents at positions 2 and 7 were prepared, and their absorption and emission spectra were studied. In particular, the synthesis route to 9-diazofluorenes with 2…
View article: MINFLUX fluorescence nanoscopy in biological tissue
MINFLUX fluorescence nanoscopy in biological tissue Open
Optical imaging access to nanometer-level protein distributions in intact tissue is a highly sought-after goal, as it would provide visualization in physiologically relevant contexts. Under the unfavorable signal-to-background conditions o…
View article: Direct optical measurement of intramolecular distances with angstrom precision
Direct optical measurement of intramolecular distances with angstrom precision Open
Optical investigations of nanometer distances between proteins, their subunits, or other biomolecules have been the exclusive prerogative of Förster resonance energy transfer (FRET) microscopy for decades. In this work, we show that MINFLU…
MINFLUX reveals dynein stepping in live neurons Open
Dynein is the primary molecular motor responsible for retrograde intracellular transport of a variety of cargoes, performing successive nanometer-sized steps within milliseconds. Due to the limited spatiotemporal precision of established m…
View article: β-Galactosidase- and Photo-Activatable Fluorescent Probes for Protein Labeling and Super-Resolution STED Microscopy in Living Cells
β-Galactosidase- and Photo-Activatable Fluorescent Probes for Protein Labeling and Super-Resolution STED Microscopy in Living Cells Open
We report on the synthesis of two fluorescent probes which can be activated by β-Galactosidase (β-Gal) enzymes and/or light. The probes contained 2-nitro-4-oxybenzyl and 3-nitro-4-oxybenzyl fragments, with β-Gal residues linked to C-4. We …
View article: MINFLUX fluorescence nanoscopy in biological tissue
MINFLUX fluorescence nanoscopy in biological tissue Open
Optical imaging access to nanometer-level protein distributions in intact tissue is a highly sought-after goal, as it would provide visualization in physiologically relevant contexts. Under the unfavorable signal-to-background conditions o…
Supramolecular Complex of Cucurbit[7]uril with Diketopyrrolopyrole Dye: Fluorescence Boost, Biolabeling and Optical Microscopy Open
New photostable and bright supramolecular complexes based on cucurbit[7]uril (CB7) host and diketopyrrolopyrole (DPP) guest dyes having two positively charged 4‐(trimethylammonio)phenyl groups were prepared and characterized. The dye core …
Supramolecular Complex of Cucurbit[7]uril with Diketopyrrolopyrole Dye: Fluorescence Boost, Biolabeling and Optical Microscopy Open
New photostable and bright supramolecular complexes based on cucurbit[7]uril (CB7) host and diketopyrrolopyrole (DPP) guest dyes having two positively charged 4‐(trimethylammonio)phenyl groups were prepared and characterized. The dye core …
Uncovering kinesin dynamics in neurites with MINFLUX Open
Neurons grow neurites of several tens of micrometers in length, necessitating active transport from the cell body by motor proteins. By tracking fluorophores as minimally invasive labels, MINFLUX is able to quantify the motion of those pro…
MINFLUX Reveals Dynein Stepping in Live Neurons Open
Dynein is the primary molecular motor responsible for retrograde intracellular transport of a variety of cargoes, performing successive nanometer-sized steps within milliseconds. Due to the limited spatiotemporal precision of established m…
Photoactivatable Xanthone (PaX) Dyes Enable Quantitative, Dual Color, and Live‐Cell MINFLUX Nanoscopy Open
The single‐molecule localization concept MINFLUX has triggered a reevaluation of the features of fluorophores for attaining nanometer‐scale resolution. MINFLUX nanoscopy benefits from temporally controlled fluorescence (“on”/“off”) photosw…
Dual-color MINFLUX: Kinesin-1 takes Chassé-Inchworm steps Open
Despite tremendous efforts using various techniques, many central questions regarding the walking mechanism of the ATP-driven motor protein kinesin-1 remained contradictory and puzzling. Still, it is widely believed that kinesin-1 walks ha…
4Pi MINFLUX arrangement maximizes spatio-temporal localization precision of fluorescence emitter Open
We introduce MINFLUX localization with interferometric illumination through opposing objective lenses for maximizing the attainable precision in 3D-localization of single inelastic scatterers, such as fluorophores. Our 4Pi optical configur…
Diffraction minima resolve point scatterers at tiny fractions (1/80) of the wavelength Open
Discerning two or more identical and constantly scattering point sources using freely propagating waves is thought to be limited by diffraction. Here we show both theoretically and experimentally that by employing a diffraction minimum rat…
Synthesis of Thioxanthone 10,10-Dioxides and Sulfone-Fluoresceins via Pd-Catalyzed Sulfonylative Homocoupling Open
Our report describes the facile and scalable preparation of 9H-thioxanthen-9-one 10,10-dioxides via Pd-catalyzed sulfonylative homocoupling of the appropriately substituted benzophenones. This transformation provides a straightforward rout…
Uncovering kinesin dynamics in neurites with MINFLUX Open
Neurons grow neurites of several tens of micrometers in length, necessitating active transport from the cell body by motor proteins. By tracking fluorophores as minimally invasive labels, MINFLUX is able to quantify the motion of those pro…
4Pi MINFLUX arrangement maximizes spatio-temporal localization precision of fluorescence emitter Open
We introduce MINFLUX localization with interferometric illumination through opposing objective lenses for maximizing the attainable precision in 3D-localization of single inelastic scatterers, such as fluorophores. Our 4Pi optical configur…
Synthesis of thioxanthone 10,10-dioxides and sulfone-fluoresceins via Pd-catalyzed sulfonylative homocoupling Open
Our report describes the facile and scalable preparation of 9H-thioxanthen-9-one 10,10-dioxides via Pd-catalyzed sulfonylative homocoupling of the appropriately substituted benzophenones. This transformation provides a straightforward entr…
MINSTED tracking of single biomolecules Open
We show that MINSTED localization, a method whereby the position of a fluorophore is identified with precisely controlled beams of a STED microscope, tracks fluorophores and hence labeled biomolecules with nanometer/millisecond spatio-temp…
View article: Photoactivatable Carbo‐ and Silicon‐Rhodamines and Their Application in MINFLUX Nanoscopy
Photoactivatable Carbo‐ and Silicon‐Rhodamines and Their Application in MINFLUX Nanoscopy Open
New photoactivatable fluorescent dyes (rhodamine, carbo‐ and silicon‐rhodamines [SiR]) with emission ranging from green to far red have been prepared, and their photophysical properties studied. The photocleavable 2‐nitrobenzyloxycarbonyl …