EDITORIAL: "When all those around me are drunk, I alone am sober," lamented an ancient Chinese poet on fighting a lone and helpless cause. In the world of science a few decades ago, there was also a lone researcher who pursued the field of optical resolution despite suspicions and derisions. Unlike those who made their names young, this scientist only succeeded thanks to his own perseverance. He developed the 4Pi microscope and successfully increased the vertical resolution of traditional optical microscopes by 3-7 times. Once he sold the patent to a company, he invested the little personal money gained from the patent in breaking the Abbe diffraction limit. Despite all his hard work, his papers kept getting rejected by respected journals, and his work was subjected to mistrust and even criticism. This scientist is Stefan Hell, someone who is never afraid of difficulties and ever determined to forge ahead.Hell was awarded the Nobel Prize in Chemistry in 2014 for being the first to propose and demonstrate that the optical diffraction limit can be broken, and for successfully developing the STED super-resolution fluorescence microscope. Later Hell and his colleagues proposed and commercialized MINFLUX, which brought optical microscopy technology to the three-dimensional single-nanometer scale, opening the "post-superresolution era".Frank and sincere, persistent and humble, boldly innovative and resolute, Hell is a scientist, an entrepreneur, a mentor, and an everyday man who enjoys life.In this interview, we will reacquaint ourselves with the Nobel laureate, Stefan Hell.
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http://dx.doi.org/10.1038/s41377-022-01034-w | DOI Listing |
Proc Natl Acad Sci U S A
December 2024
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Göttingen 37077, Germany.
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 of increased absorption and scattering of the excitation and fluorescence light in the complex tissue sample, superresolution fluorescence microscopy methods are severely challenged in attaining precise localization of molecules. We reasoned that the typical use of a confocal detection pinhole in MINFLUX nanoscopy, suppressing background and providing optical sectioning, should facilitate the detection and resolution of single fluorophores even amid scattering and optically challenging tissue environments.
View Article and Find Full Text PDFScience
October 2024
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Göttingen 37077, Germany.
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 MINFLUX fluorescence nanoscopy measures intramolecular distances down to 1 nanometer-and in planar projections down to 1 angstrom-directly, linearly, and with angstrom precision. Our method was validated by quantifying well-characterized 1- to 10-nanometer distances in polypeptides and proteins.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
September 2024
Department of Optical Nanoscopy, Max Planck Institute for Medical Research, Heidelberg 69120, Germany.
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 methods for molecular tracking, current knowledge of dynein stepping is essentially limited to slowed-down measurements in vitro. Here, we use MINFLUX fluorophore localization to directly track CRISPR/Cas9-tagged endogenous dynein with nanometer/millisecond precision in living primary neurons.
View Article and Find Full Text PDFNat Commun
August 2024
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany.
Activation of caged fluorophores in microscopy has mostly relied on the absorption of a single ultraviolet (UV) photon of ≲400 nm wavelength or on the simultaneous absorption of two near-infrared (NIR) photons >700 nm. Here, we show that two green photons (515 nm) can substitute for a single photon (~260 nm) to activate popular silicon-rhodamine (Si-R) dyes. Activation in the green range eliminates the chromatic aberrations that plague activation by UV or NIR light.
View Article and Find Full Text PDFMolecules
July 2024
Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences (MPI-NAT), Am Fassberg 11, 37077 Göttingen, Germany.
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 performed the enzymatic and photoactivation of the probes in a cuvette and compared them, prior to the labeling of fusion protein in live cells with overexpressed β-galactosidase.
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