A5082545051- Advanced Fluorescence Microscopy Techniques
- Advanced Electron Microscopy Techniques and Applications
- Cellular transport and secretion
- Force Microscopy Techniques and Applications
- Cell Image Analysis Techniques
- Lipid Membrane Structure and Behavior
- RNA and protein synthesis mechanisms
- Near-Field Optical Microscopy
- Integrated Circuits and Semiconductor Failure Analysis
- Neurobiology and Insect Physiology Research
- Cardiomyopathy and Myosin Studies
- RNA Research and Splicing
- Photosynthetic Processes and Mechanisms
- Mechanical and Optical Resonators
- Bacterial Genetics and Biotechnology
- RNA modifications and cancer
- Cell death mechanisms and regulation
- Chemical Synthesis and Analysis
- Optical Coherence Tomography Applications
- CRISPR and Genetic Engineering
- Topological and Geometric Data Analysis
- Cellular Mechanics and Interactions
- Biochemical and Structural Characterization
- Nicotinic Acetylcholine Receptors Study
- Protein Structure and Dynamics
University of Geneva
2018-2023
European Molecular Biology Laboratory
2014-2023
Sanofi (Germany)
2023
European Molecular Biology Laboratory
2013-2022
Sanofi (France)
2022
NCCR Chemical Biology - Visualisation and Control of Biological Processes Using Chemistry
2020-2021
European Bioinformatics Institute
2018-2019
European Molecular Biology Organization
2014-2019
ORCID
2015
Max Planck Institute for Developmental Biology
2011-2013
An open-source, real-time fitter for 3D single-molecule localization microscopy uses experimental point spread functions. This enables accurate super-resolution on standard microscopes without dedicated optics. We present a using functions (PSFs) that achieves minimal uncertainty in any microscope and is compatible with PSF engineering approach. used this method to image cellular structures attained unprecedented quality astigmatic PSFs. The compensates most optical aberrations makes broadly...
Highlights•High-throughput superresolution imaging of 23 proteins at thousands endocytic sites•Endocytic organize into nanoscale radial zones according to their function•A circular WASP nano-template patterns actin filament nucleation•The is crucial for high efficiency endocytosisSummaryClathrin-mediated endocytosis an essential cellular function in all eukaryotes that driven by a self-assembled macromolecular machine over 50 different tens hundreds copies. How these are organized produce...
BAX and BAK are key apoptosis regulators that mediate the decisive step of mitochondrial outer membrane permeabilization. However, mechanism by which they assemble apoptotic pore remains obscure. Here, we report present distinct oligomerization properties, with organizing into smaller structures faster kinetics than BAX. recruits accelerates assembly oligomers continue to grow during apoptosis. As a result, regulate each other as co-assemble same pores, visualize. The relative availability...
Article13 January 2022Open Access Source DataTransparent process DRP1 interacts directly with BAX to induce its activation and apoptosis Andreas Jenner orcid.org/0000-0002-6388-3053 Institute for Genetics, CECAD, University of Cologne, Germany Interfaculty Biochemistry, Tübingen, Contribution: Formal analysis (equal), Investigation Visualization Methodology Writing - review & editing (equal) Search more papers by this author Aida Peña-Blanco Raquel Salvador-Gallego...
Clathrin-mediated endocytosis is an essential process that forms vesicles from the plasma membrane. Although most of protein components endocytic machinery have been thoroughly characterized, their organization at site poorly understood. We developed a fluorescence microscopy method to track average positions yeast proteins in relation each other with time precision below 1 s and spatial ∼10 nm. With these data, integrated shapes membrane intermediates superresolution imaging, we could...
Eukaryotic cells use clathrin-mediated endocytosis to take up a large range of extracellular cargo. During endocytosis, clathrin coat forms on the plasma membrane, but it remains controversial when and how is remodeled into spherical vesicle. Here, we 3D superresolution microscopy determine precise geometry at numbers endocytic sites. Through pseudo-temporal sorting, average trajectory remodeling during endocytosis. We find that coats assemble first flat membranes 50% area before they become...
We present a fundamentally new approach to 3D superresolution microscopy based on the principle of surface-generated fluorescence. This near-field fluorescence is strongly dependent distance fluorophores from coverslip and can therefore be used estimate their axial positions. established robust simple implementation supercritical angle detection for single-molecule localization microscopy, calibrated it using fluorescent bead samples, validated method with DNA origami tetrahedra,...
Localization microscopy data is represented by a set of spatial coordinates, each corresponding to single detection, that form point cloud. This can be analyzed either rendering an image from these or analyzing the cloud directly. Analysis this type has focused on clustering detections into distinct groups which produces measurements such as cluster area, but limited capacity quantify complex molecular organization and nano-structure.We present segmentation protocol which, through...
Three-dimensional single molecule localization microscopy relies on the fitting of individual molecules with a point spread function (PSF) model. The reconstructed images often show local squeezing or expansion in z. A common cause is depth-induced aberrations conjunction an imperfect PSF model calibrated from beads coverslip, resulting mismatch between measured and real PSF. Here, we developed strategy for accurate z-localization which use fitting, determine errors correct them...
Quantitative data analysis is important for any single-molecule localization microscopy (SMLM) workflow to extract biological insights from the coordinates of single fluorophores. However, current approaches are restricted simple geometries or require identical structures. Here, we present LocMoFit (Localization Model Fit), an open-source framework fit arbitrary model coordinates. It extracts meaningful parameters individual structures and can select most suitable model. In addition...
Essential for eukaryotic organisms, multiple copies of the exocyst complex tether each secretory vesicle to plasma membrane (PM) in constitutive exocytosis. The higher-order structure (ExHOS) that coordinates action these exocysts remains unexplored. We integrated particle-tracking, super-resolution microscopy and cryo-electron tomography a model time-resolves continuum conformational landscape ExHOS functionally annotates its different conformations. found 7 form flexible ring-shaped...
Clathrin-mediated endocytosis in budding yeast requires the formation of a dynamic actin network that produces force to invaginate plasma membrane against intracellular turgor pressure. The type-I myosins Myo3 and Myo5 are important for endocytic reshaping, but mechanistic details their function remain scarce. Here, we studied during using quantitative live-cell imaging genetic perturbations. We show promote, dose-dependent way, growth expansion network, which controls speed coat...
Synaptic membrane-remodeling events such as endocytosis require force-generating actin assembly. The endocytic machinery that regulates these and membrane dynamics localizes at high concentrations to large areas of the presynaptic membrane, but assembly productive are far more restricted in space time. Here we describe a mechanism whereby autoinhibition clamps limit discrete functional events. We found collective interactions between Drosophila proteins Nwk/FCHSD2, Dap160/intersectin, WASp...
Seeing the big picture: Asymmetric macromolecular complexes that are NMR active in only a subset of their subunits can be prepared, thus decreasing spectral complexity. For hetero heptameric LSm1–7 and LSm2–8 rings spectra individual complete complex obtained, showing conserved RNA binding site. This LEGO-NMR technique makes large asymmetric accessible to detailed spectroscopic studies. spectroscopy is unique as it provides means study bio-molecules with atomic resolution near natural...
A broad variety of biomolecules is industrially produced in bacteria and yeasts. These microbial expression hosts can be optimized through genetic engineering using CRISPR tools. Here, we designed characterized such a modular genome editing system based on the Cas12a-like RNA-guided nuclease MAD7 Escherichia coli. This enables efficient generation single nucleotide polymorphisms (SNPs) or gene deletions directly used with donor DNA from benchtop assembly to increase throughput. We combined...
Abstract Quantitative fluorescence and superresolution microscopy are often limited by insufficient data quality or artifacts. In this context, it is essential to have biologically relevant control samples benchmark optimize the of microscopes, labels imaging conditions. Here we exploit stereotypic arrangement proteins in nuclear pore complex as situ reference structures characterize performance a variety modalities. We created four genome edited cell lines which endogenously labeled...
Abstract Eukaryotic cells use clathrin-mediated endocytosis to take up a large range of extracellular cargos. During endocytosis, clathrin coat forms on the plasma membrane, but it remains controversial when and how is remodeled into spherical vesicle. Here, we 3D superresolution microscopy determine precise geometry at numbers endocytic sites. Through pseudo-temporal sorting, average trajectory remodeling during endocytosis. We find that coats assemble first flat membranes 50% area, before...
Clathrin-mediated endocytosis is an essential cellular function in all eukaryotes that driven by a self-assembled macromolecular machine of over 50 different proteins tens to hundreds copies. How these are organized produce endocytic vesicles with high precision and efficiency not understood. Here, we developed high-throughput superresolution microscopy reconstruct the nanoscale structural organization 23 from 100,000 sites yeast. We found assemble radially-ordered recruitment according...