A5029490828- Advanced Fluorescence Microscopy Techniques
- Lipid Membrane Structure and Behavior
- Cell Image Analysis Techniques
- RNA Research and Splicing
- Force Microscopy Techniques and Applications
- Peroxisome Proliferator-Activated Receptors
- Neuroscience and Neuropharmacology Research
- T-cell and B-cell Immunology
- interferon and immune responses
- Genomics and Chromatin Dynamics
- Single-cell and spatial transcriptomics
- HIV Research and Treatment
- Cellular Mechanics and Interactions
- Spectroscopy and Quantum Chemical Studies
- Animal testing and alternatives
- Biosimilars and Bioanalytical Methods
- Immunotherapy and Immune Responses
- Spectroscopy Techniques in Biomedical and Chemical Research
- Ion channel regulation and function
- Optical Imaging and Spectroscopy Techniques
- Genomics and Phylogenetic Studies
- Chromosomal and Genetic Variations
- PARP inhibition in cancer therapy
- RNA modifications and cancer
- Calcium signaling and nucleotide metabolism
University of Oxford
2015-2024
MRC Weatherall Institute of Molecular Medicine
2015-2024
John Radcliffe Hospital
2015-2024
MRC Human Immunology Unit
2015-2023
Medical Research Council
2020-2022
Imaging Center
2015-2019
Institute for Molecular Medicine
2017
University College London
2008-2015
Landscape Research Group
2014
Inserm
2008
First-in-class clinical trial of IL-33 blockade shows inhibition neutrophil migration into human skin and modulates atopic dermatitis symptoms.
Activating T cells reorganize their cortical actin to form a ramified transportation network beneath the immunological synapse.
The three-dimensional (3D) organization of chromosomes can be probed using methods like Capture-C. However, it is unclear how such population-level data relate to the within a single cell, and mechanisms leading observed interactions are still largely obscure. We present polymer modeling scheme based on assumption that chromosome architecture maintained by protein bridges, which form chromatin loops. To test model, we perform FISH experiments compare with Capture-C data. Starting merely from...
Diffusion and interaction dynamics of molecules at the plasma membrane play an important role in cellular signaling are suggested to be strongly associated with actin cytoskeleton. Here we use superresolution STED microscopy combined fluorescence correlation spectroscopy (STED-FCS) access compare diffusion characteristics fluorescent lipid analogues GPI-anchored proteins (GPI-APs) live-cell cytoskeleton-free, cell-derived giant vesicles (GPMVs). Hindered phospholipids sphingolipids is...
The β-subunits of voltage-gated calcium channels regulate their functional expression and properties. Two mechanisms have been proposed for this, an effect on gating enhancement expression. With respect to the expression, suggested enhance trafficking by masking unidentified endoplasmic reticulum (ER) retention signal. Here we investigated whether, how, affect level Ca(V)2.2 within somata neurites cultured sympathetic neurons. We used YFP-Ca(V)2.2 containing a mutation (W391A), that prevents...
Abstract Physicochemical properties of the plasma membrane have been shown to play an important role in cellular functionality. Among those properties, molecular order lipids, or lipid packing, is high importance. Changes packing are believed compartmentalize signaling by initiating coalescence and conformational changes proteins. A common way infer using membrane‐embedded polarity‐sensitive dyes, whose emission spectrum dependent on immediate environment. Here, we report improved...
Measuring small forces is a major challenge in cell biology. Here we improve the spatial resolution and accuracy of force reconstruction well-established technique traction microscopy (TFM) using STED microscopy. The increased STED-TFM (STFM) allows greater than 5-fold higher sampling generated by conventional TFM, accessing nano instead micron scale. This improvement highlighted computer simulations an activating RBL model system.
Human immunodeficiency virus type 1 (HIV-1) assembles as immature particles, which require the proteolytic cleavage of structural polyprotein Gag and clustering envelope glycoprotein Env for infectivity. The details mechanisms underlying remain unknown. Here, we determine molecular dynamics on surface individual HIV-1 particles using scanning fluorescence correlation spectroscopy a super-resolution STED microscope. We find that undergoes maturation-induced increase in mobility,...
Integrins are the core constituents of cell-matrix adhesion complexes such as focal adhesions (FAs) and play key roles in physiology disease. fluctuate between active inactive conformations, yet whether activity state influences spatial organization integrins within FAs has remained unclear. In this study, we address question also ask integrin may be regulated either independently for each molecule or through locally coordinated mechanisms. We used two distinct superresolution microscopy...
Cholesterol (Chol) is a crucial component of cellular membranes, but knowledge its intracellular dynamics scarce. Thus, it utmost interest to develop tools for visualization Chol organization and in cells tissues. For this purpose, many studies make use fluorescently labeled analogs. Unfortunately, the introduction label may influence characteristics analog, such as localization, interaction, trafficking cells; hence, important get bias. In report, we compared different fluorescent lipid...
Although plasma leakage is the hallmark of severe dengue infections, factors that cause increased vascular permeability have not been identified. As platelet activating factor (PAF) associated with an increase in other diseases, we set out to investigate its role acute infection.PAF levels were initially assessed 25 patients infection determine if they dengue. For investigation kinetics PAF, serial PAF values 36 patients. The effect serum on tight junction protein ZO-1 was determined by...
Heterogeneous diffusion dynamics of molecules play an important role in many cellular signaling events, such as lipids plasma membrane bioactivity. However, these can often only be visualized by single-molecule and super-resolution optical microscopy techniques. Using fluorescence lifetime correlation spectroscopy (FLCS, extension spectroscopy, FCS) on a stimulated emission depletion (STED) microscope, we here extend previous observations nanoscale lipid the living mammalian cells. STED-FLCS...
Membrane-associated events during peroxisomal protein import processes play an essential role in peroxisome functionality. Many details of these are not known due to missing spatial resolution technologies capable investigating peroxisomes directly the cell. Here, we present use super-resolution optical stimulated emission depletion microscopy investigate with sub-60-nm heterogeneous organization proteins PEX5, PEX14, and PEX11 around actively importing peroxisomes, showing distinct...
Abstract Self-interacting chromatin domains encompass genes and their cis -regulatory elements; however, the three-dimensional form a domain takes, whether this relies on enhancer–promoter interactions, processes necessary to mediate formation maintenance of such domains, remain unclear. To examine these questions, here we use combination high-resolution chromosome conformation capture, non-denaturing fluorescence in situ hybridisation super-resolution imaging study 70 kb encompassing mouse...
The lateral organization of molecules in the cellular plasma membrane plays an important role signaling. A critical parameter for molecular is how lipids are packed. Polarity-sensitive dyes powerful tools to characterize such lipid order, employing, example, confocal and two-photon microscopy. investigation potential nanodomains, however, requires use superresolution Here, we test performance polarity-sensitive Di-4-ANEPPDHQ, Di-4-AN(F)EPPTEA, NR12S stimulated emission depletion Measurements...
Abstract Motivation: Fluorescence Correlation Spectroscopy (FCS) is a popular tool for measuring molecular mobility and how relates to interaction dynamics bioactivity in living cells. The FCS technique has been significantly advanced by its combination with super-resolution STED microscopy (STED-FCS). Specifically, the use of gated detection shown great potential enhancing STED-FCS, but also created demand software which efficient implements latest algorithms. Prior this study, no open...
The antigen-presenting molecule MR1 presents riboflavin-based metabolites to Mucosal-Associated Invariant T (MAIT) cells. While egress the cell surface is ligand-dependent, ability of small-molecule ligands impact on cellular trafficking remains unknown. Arising from an in silico screen ligand-binding pocket, we identify one ligand, 3-([2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl]formamido)propanoic acid, DB28, as well analog, methyl...
Recent years have seen the development of multiple technologies to investigate, with great spatial and temporal resolution, dynamics lipids in cellular model membranes. One these approaches is combination far-field super-resolution stimulated-emission-depletion (STED) microscopy fluorescence correlation spectroscopy (FCS). STED-FCS combines diffraction-unlimited resolution STED statistical accuracy FCS determine sub-millisecond-fast molecular single-molecule sensitivity. A unique advantage...
Expansion microscopy (ExM) has been successfully used to improve the spatial resolution when imaging tissues by optical microscopy. In ExM, proteins of a fixed sample are crosslinked swellable acrylamide gel, which expands incubated in water. Therefore, ExM allows enlarged subcellular structures be resolved that would otherwise hidden standard confocal Herein, we aim validate for study peroxisomes, mitochondria, nuclei and plasma membrane. Upon comparison expansion factors these cellular...
Antibodies against the Membrane-Proximal External Region (MPER) of Env gp41 subunit neutralize HIV-1 with exceptional breadth and potency. Due to lack knowledge on MPER native structure accessibility, different exclusive models have been proposed for molecular mechanism recognition by broadly neutralizing antibodies. Here, accessibility antibodies single virions has addressed through STED microscopy. imaging fluorescently labeled Fabs reveals a common pattern targeting or surface gp120. In...
Object detection networks are high-performance algorithms famously applied to the task of identifying and localizing objects in photography images. We demonstrate their application for classification localization cells fluorescence microscopy by benchmarking four leading object across multiple challenging 2D datasets. Furthermore we develop an algorithm that can localize image 3D, close real time, at microscope using widely available inexpensive hardware. Furthermore, exploit fast processing...
Kinase oxidation is a critical signaling mechanism through which changes in the intracellular redox state alter cardiac function. In myocardium, PKARIα (type-1 protein kinase A) can be reversibly oxidized, forming interprotein disulfide bonds holoenzyme complex. However, effect of formation on downstream heart, particularly under states oxidative stress such as ischemia and reperfusion (I/R), remains unexplored.Atrial tissue obtained from patients before after cardiopulmonary bypass left...
We report the development and testing of software called QuantiFly: an automated tool to quantify Drosophila egg laying. Many laboratories count eggs as a marker fitness. The existing method requires laboratory researchers manually while looking down microscope. This technique is both time-consuming tedious, especially when experiments require daily counts hundreds vials. basis QuantiFly algorithm which applies improves upon advanced pattern recognition machine-learning routine. accuracy...