A5055947936- Advanced Fluorescence Microscopy Techniques
- Advanced Electron Microscopy Techniques and Applications
- Near-Field Optical Microscopy
- Spectroscopy Techniques in Biomedical and Chemical Research
- Cell Image Analysis Techniques
- Thermography and Photoacoustic Techniques
- Gold and Silver Nanoparticles Synthesis and Applications
- Plasmonic and Surface Plasmon Research
- Photosynthetic Processes and Mechanisms
- Photoacoustic and Ultrasonic Imaging
- Advanced Optical Sensing Technologies
- Optical Coherence Tomography Applications
- Adhesion, Friction, and Surface Interactions
- Advanced X-ray Imaging Techniques
- Nerve injury and regeneration
- Electron Spin Resonance Studies
- Thermal properties of materials
- Metabolism and Genetic Disorders
- Photonic and Optical Devices
- Molecular Biology Techniques and Applications
- Toxin Mechanisms and Immunotoxins
- Neuroscience and Neural Engineering
- ATP Synthase and ATPases Research
- Advanced biosensing and bioanalysis techniques
- Force Microscopy Techniques and Applications
Research Institute of Molecular Pathology
2021-2023
Vienna Biocenter
2023
Max Planck Institute for Multidisciplinary Sciences
2022
Max Planck Institute for Biophysical Chemistry
2012-2020
University of Buenos Aires
2007-2009
Superresolution imaging in sharper focus An optical microscope cannot distinguish objects separated by less than half the wavelength of light. techniques have broken this “diffraction limit” and provided exciting new insights into cell biology. Still, such hit a limit at resolution about 10 nm. Balzarotti et al. describe another way localizing single molecules called MINFLUX (see Perspective Xiao Ha). As photoactivated localization microscopy stochastic reconstruction microscopy,...
Mitochondrial function is critically dependent on the folding of mitochondrial inner membrane into cristae; indeed, numerous human diseases are associated with aberrant crista morphologies. With MICOS complex, OPA1 and F1 Fo -ATP synthase, key players cristae biogenesis have been identified, yet their interplay poorly understood. Harnessing super-resolution light 3D electron microscopy, we dissect roles these proteins in formation mitochondria. We individually disrupted genes all seven...
Significance Popular localization of single molecules through calculating the centroid diffraction pattern produced by molecular fluorescence on a camera is typically limited to spatiotemporal resolutions >10 nm per milliseconds. By requiring at least 10–100 times fewer detected photons and being free bias due orientation, concept called MINFLUX propels tracking hitherto-unachievable regime single-digit nanometer precision within substantially less than millisecond. Our experiments herald...
Significance In vertebrates, the action potential travels along myelin-coated electrically isolated axons and is regenerated at nodes of Ranvier, which lack myelination are characterized by a tight interaction between axon glial cells. Specific sets proteins enriched in each region nodes. Thanks to its subdiffraction resolution, stimulated emission depletion (STED) microscopy here uncovers organization 12 these nanoscale. The superresolved imaging reveals an extremely fine interplay...
Significance Superresolution fluorescence microscopy (nanoscopy) has enabled the study of protein distributions in cellular organelles, albeit only down to a 3D resolution 20 40 nm. By targeting single emitters with doughnut-shaped excitation beam, recently introduced MINFLUX nanoscopy provides single-digit nanometer localizations fluorescent labels. We show an application multicolor imaging densely packed labeled proteins, specifically subunits MICOS complex, large complex within...
We show that RESOLFT fluorescence nanoscopy, a low light level scanning superresolution technique employing reversibly switchable fluorescent proteins (rsFPs), is capable of dual-channel live-cell imaging virtually free chromatic errors and temporal offsets. This accomplished using rsEGFP Dronpa, two rsFPs having similar spectra but different kinetics switching emission. Our approach demonstrated by protein distributions dynamics in living neurons neuronal tissues.
Commonly, in stimulated emission depletion (STED) fluorescence nanoscopy, light of a wavelength located at the red tail spectrum dye is used to shrink effective fluorophore excitation volume and thus obtain images with sub diffraction resolution.Here, we demonstrate that continuous wave (CW) STED nanoscopy feasible using wavelengths maximum, where cross section for up 10-fold larger than tail.As result, imaging becomes possible equally lower beam power.Besides, fluorophores have been...
Michel Orrit opened a discussion of the paper by Paola Borri: You showed an interpretation spectral response gold nanoparticles in terms shift and broadening surface plasmon band. This empirical description is model-independent. Did your data fit with only sh
The ultimate goal of biological superresolution fluorescence microscopy is to provide three-dimensional resolution at the size scale a fluorescent marker. Here, we show that, by localizing individual switchable fluorophores with probing doughnut-shaped excitation beam, MINFLUX nanoscopy provides 1–3 nanometer in fixed and living cells. This progress has been facilitated approaching each fluorophore iteratively doughnut minimum, making essentially uniform isotropic over scalable fields view....
We present a novel noncontact, photothermal technique, based on the focus error signal of commercial CD pickup head that allows direct determination absorption in thin films. Combined with extinction methods, this technique yields scattering contribution to losses. Surface plasmon polaritons are excited using Kretschmann configuration Au films varying thickness. By measuring and simultaneously, it is shown dielectric constants thickness retrieval leads inconsistencies if model does not...
Abstract Beams of light that feature an intensity zero are essential to a variety optical microscopy methods. Super-resolution techniques like STED and RESOLFT, together with localization strategies MINFLUX MINSTED, rely on accurate fast displacements such beams their zeros. Extending these methods the third dimension requires axial deflection, which, in contrast lateral remains technologically challenging microsecond scale. Here, we present general-purpose beam-shaping polarization...
Abstract Prakash and Curd provide a re-analysis 1 of individual datasets taken from our report 2 demonstrating MINFLUX 3D imaging in cells. Their evaluation confirms the unique localization precision provided by 2,3 featuring standard deviation σ = 1-3 nm. We appreciate their confirmation also welcome opportunity to clarify remaining points. The hitherto almost unconceivable attained is likely hold key an all-optical dynamical structural biology.
Maxime Dahan opened the discussion of paper by William Moerner: You present a method for 3D localization over more than 10 microns. What is trade-off between axial depth and accuracy? Moerner replied: It important to note that since new PSFs ar
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