A5005209379- Platelet Disorders and Treatments
- Advanced Fluorescence Microscopy Techniques
- Blood properties and coagulation
- Cellular Mechanics and Interactions
- Cell Adhesion Molecules Research
- Advanced Electron Microscopy Techniques and Applications
- Antiplatelet Therapy and Cardiovascular Diseases
- Neuroscience and Neural Engineering
- Lipid Membrane Structure and Behavior
- Cell Image Analysis Techniques
- Blood groups and transfusion
- Force Microscopy Techniques and Applications
- Genetics, Aging, and Longevity in Model Organisms
- Near-Field Optical Microscopy
- Microfluidic and Bio-sensing Technologies
- Electrospun Nanofibers in Biomedical Applications
- Blood Coagulation and Thrombosis Mechanisms
- Complement system in diseases
- Spaceflight effects on biology
- Monoclonal and Polyclonal Antibodies Research
- Venous Thromboembolism Diagnosis and Management
- Integrated Circuits and Semiconductor Failure Analysis
- Polymer Surface Interaction Studies
- 3D Printing in Biomedical Research
- Advanced Biosensing Techniques and Applications
Royal College of Surgeons in Ireland
2017-2024
Inserm
2023
Université de Montpellier
2023
Centre National de la Recherche Scientifique
2023
Centre de Biologie Structurale
2023
ETH Zurich
2011-2022
Charles Humbert 8
2017
Department of Medical Sciences
2017
École Polytechnique Fédérale de Lausanne
2010
Ludwig-Maximilians-Universität München
2009-2010
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...
Abstract Dysfunctional extracellular matrices (ECM) contribute to aging and disease. Repairing dysfunctional ECM could potentially prevent age-related pathologies. Interventions promoting longevity also impact gene expression. However, the role of composition changes in healthy remains unclear. Here we perform proteomics in-vivo monitoring systematically investigate (matreotype) during C. elegans revealing three distinct collagen dynamics. Longevity interventions slow stiffening prolong...
Quantifying cellular forces relies on accurate calibrations of the sensor stiffness. Neglecting deformations elastic substrates to which pillars are anchored systematically overestimates applied (up 40%). A correction factor considering substrate warping is derived analytically and verified experimentally. The scales with dimensionless pillar aspect ratio. This has significant implications when designing arrays or comparing absolute measured different geometries during cell spreading,...
Many nucleic acid stains show a strong fluorescence enhancement upon binding to double-stranded DNA. Here we exploit this property perform superresolution microscopy based on the localization of individual events. The dynamic labeling scheme and optimization fluorophore brightness yielded resolution ∼14 nm (fwhm) spatial sampling 1/nm. We illustrate our approach with two different DNA-binding dyes apply it visualize organization bacterial chromosome in fixed Escherichia coli cells. In...
According to the Chinese yin-yang concept, seemingly opposing forces give rise and respond each other. Opposing forces, whether passive or active, are also at work when cells adhere a substrate extracellular matrix, sense environmental properties, finally them. In this review, we describe molecular elements inside outside of cell that establish labile physical connections, how regulate their interplay, namely formation, reinforcement, breakage, reconfiguration these elements. What locally...
Abstract Fibronectin fibrils within the extracellular matrix play central roles in physiological and pathological processes, yet many structural details about their hierarchical molecular assembly remain unknown. Here we combine site-specific protein labelling with single-molecule localization by stepwise photobleaching or direct stochastic optical reconstruction microscopy (dSTORM), determine relative positions of various labelled sites native fibrils. Single end-labelled fibronectin...
Abstract Dendritic cells play a key role in processing and presenting antigens to naïve T prime adaptive immunity. Circadian rhythms are known regulate many aspects of immunity; however, the circadian dendritic cell function is still unclear. Here, we show greater responses when mice immunised middle their rest versus active phase. We find rhythm antigen that correlates with both mitochondrial morphology metabolism, dependent on molecular clock gene, Bmal1 . Using Mdivi-1, compound promotes...
Reliable extracellular stimulation of neuronal activity is the prerequisite for electrical interfacing cultured networks and brain slices, as well neural implants. Safe must be achieved without damage to cells. With respect a future application highly integrated semiconductor chips, we present an electrophysiological study capacitive mammalian cells in geometry adhesion on insulated titanium dioxide/silicon electrode. We used HEK293 with overexpressed Na(V)1.4 channels neurons from rat...
It is generally expected that the kinetics of reactions inside living cells differs from situation in bulk solutions. Macromolecular crowding and specific binding interactions could change diffusion properties availability free molecules. Their impact on reaction relevant context still elusive, mainly because difficulty capturing fast vivo. This article shows spatially resolved measurements DNA hybridization single cells. HeLa were transfected with a FRET-labeled dsDNA probe by lipofection....
Specific interactions of protein side chains and lipid membranes regulate the localization, orientation, activity many peripheral proteins. Here, we introduce a modification coarse-grained MARTINI model, called 'side chain fix' (scFix), that was necessary sufficient to correctly sample dynamics β-strands in several globular When compared μs long atomistic simulations or previous experimental findings, scFix reproduced all key between well-studied PLC-δ1 pleckstrin homology domain...
Protein unfolding induced by nanoparticles (NPs) can lead to exposure of cryptic epitopes that might dictate biological identity and affect NP fate (e.g., blood circulation time, biodistribution, tumor accumulation). Here, we monitor the conformation fluorescence resonance energy transfer (FRET)-labelled fibronectin (FN) on corona-coated gold NPs. We found labelled FN proteins, which directly accessed surface, underwent more pronounced conformational changes than those associated with...
The precise spatial localization of proteins in situ by super-resolution microscopy (SRM) demands their targeted labeling. Positioning reporter molecules as close possible to the target remains a challenge primary cells or tissues from patients that cannot be easily genetically modified. Indirect immunolabeling introduces relatively large linkage errors, whereas site-specific and stoichiometric labeling antibodies relies on elaborate chemistries. In this study, we developed simple two-step...
Breast cancer results in a three- to four-fold increased risk of venous thromboembolism (VTE), which is associated with reduced patient survival. Despite this, the mechanisms underpinning breast cancer-associated thrombosis remain poorly defined. Tumor cells can trigger endothelial cell (EC) activation resulting von Willebrand factor (VWF) secretion. Importantly, elevated plasma VWF levels constitute an independent biomarker for VTE risk. Moreover, model melanoma, treatment low molecular...
Abstract The plasma multimeric glycoprotein von Willebrand factor (VWF) plays a critical role in primary hemostasis by tethering platelets to exposed collagen at sites of vascular injury. Recent studies have identified additional biological roles for VWF, and particular suggest that VWF may play an important regulating inflammatory responses. However, the molecular mechanisms through which exerts its immuno-modulatory effects remain poorly understood. In this study, we report binding...
Phagocytes, predatory cells of the immune system, continuously probe their cellular microenvironment on hunt for invaders. This requires prey recognition followed by formation physical contacts sufficiently stable pickup. Although must apply forces to pick up microbial prey, little is known about hunting behavior preceding phagocytosis because a lack appropriate technologies. To study phagocyte in which adhesive bonds holds surfaces be broken, we exploited use microrobotic probes mimic...
MYH9 -related disease patients with mutations in the contractile protein nonmuscle myosin heavy chain IIA display, among others, macrothrombocytopenia and a mild-to-moderate bleeding tendency. In this study, we used three mouse lines, each one point mutation Myh9 gene at positions 702, 1424, or 1841, to investigate mechanisms underlying increased risk. Agonist-induced activation of mutant platelets was comparable controls. However, light phosphorylation after reduced platelets, which...
Early wound healing is associated with fibroblasts assembling a provisional fibronectin-rich extracellular matrix (ECM), which subsequently remodeled and interlaced by type I collagen. This exposes to time-variant sets of matrices during different stages healing. Our goal was thus gain insight into the ECM-driven functional regulation human foreskin (HFFs) being either anchored fibronectin (Fn) or collagen-decorated matrix, in absence presence cyclic mechanical strain. While cells reoriented...
Abstract Advances in nanofabrication methods have enabled the tailoring of new strategies towards controlled production nanoparticles with attractive applications healthcare. In many cases, their characterisation remains a big challenge, particularly for small-sized functional 5 nm diameter or smaller, where current particle sizing techniques struggle to provide required sensitivity and accuracy. There is clear need development reliable approaches physico-chemical significant accuracy,...
Platelets interact with multiple adhesion proteins during thrombogenesis, yet little is known about their ability to assemble fibronectin matrix. In vitro three-dimensional superresolution microscopy complemented by biophysical and biochemical methods revealed fundamental insights into how platelet contractility drives fibrillogenesis. adhering thrombus (fibronectin fibrin) versus basement membrane components (laminin collagen IV) pull fibrils along apical underneath basal membrane,...
Abstract The initial contact with blood and its components, including plasma proteins platelets, directs the body's response to foreign materials. Natural scaffolds of extracellular matrix or fibrin contain fibrils nanoscale dimensions, but how platelets specifically respond topography architecture fibrous materials is still incompletely understood. Here, planar nanofiber are fabricated from native fibrinogen characterize morphology adherent activation markers for phosphatidylserine exposure...
Haemostatic platelet function is intimately linked to cellular mechanics and cytoskeletal morphology. How reorganizations give rise a highly contractile phenotype that necessary for clot contraction remains poorly understood. To elucidate this process in vitro, we developed morphometric screen quantify the spatial organization of actin fibres vinculin adhesion sites single spread platelets. Platelets from healthy donors predominantly adopted bipolar morphology on fibrinogen fibronectin,...