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Results: 27
Activation of Toll-like receptors nucleates assembly of the MyDDosome signaling hub.
Sarah Louise Latty, Jiro Sakai, Lee Hopkins, Brett Verstak, Teresa Paramo, Nils A Berglund, Eugenia Cammarota, Pietro Cicuta, Nicholas J Gay, Peter J Bond, David Klenerman, Clare E Bryant
Mar 23, 2018
Infection and tissue damage induces assembly of supramolecular organizing centres (SMOCs)), such as the Toll-like receptor (TLR) MyDDosome, to co-ordinate inflammatory signaling. SMOC assembly is thought to drive digital...
Hsp70 Inhibits the Nucleation and Elongation of Tau and Sequesters Tau Aggregates with High Affinity.
Franziska Kundel, Suman De, Patrick Flagmeier, Mathew H Horrocks, Magnus Kjaergaard, Sarah L Shammas, Sophie E Jackson, Christopher M Dobson, David Klenerman
Mar 13, 2018
Systematic Assessment of the Accuracy of Subunit Counting in Biomolecular Complexes Using Automated Single-Molecule Brightness Analysis.
John SH Danial, Yuri Quintana, Uris Ros, Raed Shalaby, Eleonora G Margheritis, Sabrina Chumpen Ramirez, Christian Ungermann, Ana J Garcia-Saez, Katia Cosentino
Feb 19, 2022
Analysis of single-molecule brightness allows subunit counting of high-order oligomeric biomolecular complexes. Although the theory behind the method has been extensively assessed, systematic analysis of the experimental...
Single-Molecule Light-Sheet Imaging of Suspended T Cells.
Aleks Ponjavic, James McColl, Alexander R Carr, Ana Mafalda Santos, Klara Kulenkampff, Anna Lippert, Simon J Davis, David Klenerman, Steven F Lee
Apr 11, 2019
Adaptive immune responses are initiated by triggering of the T cell receptor. Single-molecule imaging based on total internal reflection fluorescence microscopy at coverslip/basal cell interfaces is commonly used to study this...
Ultra-High Resolution 3D Imaging of Whole Cells.
Fang Huang, George Sirinakis, Edward S Allgeyer, Lena K Schroeder, Whitney C Duim, Emil B Kromann, Thomy Phan, Felix E Rivera-Molina, Jordan R Myers, Irnov Irnov, Mark Lessard, Yongdeng Zhang, Mary Ann Handel, Christine Jacobs-Wagner, C Patrick Lusk, James E Rothman, Derek Toomre, Martin J Booth, Joerg Bewersdorf
Apr 15, 2019
Fluorescence nanoscopy, or super-resolution microscopy, has become an important tool in cell biological research. However, because of its usually inferior resolution in the depth direction (50-80 nm) and rapidly deteriorating...
Live-cell imaging reveals the spatiotemporal organization of endogenous RNA polymerase II phosphorylation at a single gene.
Linda S Forero-Quintero, William Raymond, Tetsuya Handa, Matthew N Saxton, Tatsuya Morisaki, Hiroshi Kimura, Edouard Bertrand, Brian Munsky, Timothy J Stasevich
Jun 27, 2021
The carboxyl-terminal domain of RNA polymerase II (RNAP2) is phosphorylated during transcription in eukaryotic cells. While residue-specific phosphorylation has been mapped with exquisite spatial resolution along the 1D genome...
Bioorthogonal protein-DNA conjugation methods for force spectroscopy.
Accurate and stable site-specific attachment of DNA molecules to proteins is a requirement for many single-molecule force spectroscopy techniques. The most commonly used method still relies on maleimide chemistry involving...
Single-molecule visualization of DNA G-quadruplex formation in live cells.
Marco Di Antonio, Aleks Ponjavic, Antanas Radzevičius, Rohan T Ranasinghe, Marco Catalano, Xiaoyun Zhang, Jiazhen Shen, Lisa-Maria Needham, Steven F Lee, David Klenerman, Shankar Balasubramanian
Jun 23, 2020
Quantification of very low-abundant proteins in bacteria using the HaloTag and epi-fluorescence microscopy.
Alessia Lepore, Hannah Taylor, Dirk Landgraf, Burak Okumus, Sebastián Jaramillo-Riveri, Lorna McLaren, Somenath Bakshi, Johan Paulsson, M El Karoui
Aug 19, 2019
Cell biology is increasingly dependent on quantitative methods resulting in the need for microscopic labelling technologies that are highly sensitive and specific. Whilst the use of fluorescent proteins has led to major...
Single-Molecule Analysis of G Protein-Coupled Receptor Stoichiometry
How G protein-coupled receptors (GPCRs) are organized at the cell surface remains highly contentious. Single-molecule (SM) imaging is starting to inform this debate as receptor behavior can now be visualized directly, without...
A cell topography-based mechanism for ligand discrimination by the T cell receptor.
Ricardo A Fernandes, Kristina A Ganzinger, Justin C Tzou, Peter Jönsson, Steven F Lee, Matthieu Palayret, Ana Mafalda Santos, Alexander R Carr, Aleks Ponjavic, Veronica T Chang, Charlotte Macleod, B Christoffer Lagerholm, Alan E Lindsay, Omer Dushek, Andreas Tilevik, Simon J Davis, David Klenerman
May 24, 2019
T cell receptor dwell time microvilli receptor triggering single-molecule imaging Animals Antigen-Presenting Cells Host-Pathogen Interactions Humans Immunity Innate Kinetics Ligands Lymphocyte Activation Major Histocompatibility Complex Microvilli Models Theoretical Peptides Phosphorylation Receptors Antigen T-Cell Signal Transduction single molecule imaging T-Lymphocytes
Somatic mutation landscapes at single-molecule resolution.
Federico Abascal, Luke MR Harvey, Emily Mitchell, Andrew RJ Lawson, Stefanie V Lensing, Peter Ellis, Andrew JC Russell, Raul E Alcantara, Adrian Baez-Ortega, Yichen Wang, Eugene Jing Kwa, Henry Lee-Six, Alex Cagan, Tim HH Coorens, Michael Spencer Chapman, Sigurgeir Olafsson, Steven Leonard, David Jones, Heather E Machado, Megan Davies, Nina F Øbro, Krishnaa T Mahubani, Kieren Allinson, Moritz Gerstung, Kourosh Saeb-Parsy, David G Kent, Elisa Laurenti, Michael R Stratton, Raheleh Rahbari, Peter J Campbell, Robert J Osborne, Iñigo Martincorena
Mar 24, 2021
Somatic mutations drive the development of cancer and may contribute to ageing and other diseases1,2. Despite their importance, the difficulty of detecting mutations that are only present in single cells or small clones has...
Challenges of analysing stochastic gene expression in bacteria using single-cell time-lapse experiments.
Stochastic gene expression causes phenotypic heterogeneity in a population of genetically identical bacterial cells. Such non-genetic heterogeneity can have important consequences for the population fitness, and therefore cells...
Cryo-EM single-particle structure refinement and map calculation using Servalcat.
In 2020, cryo-EM single-particle analysis achieved true atomic resolution thanks to technological developments in hardware and software. The number of high-resolution reconstructions continues to grow, increasing the importance...
Oligomer Diversity during the Aggregation of the Repeat Region of Tau.
Magnus Kjaergaard, Alexander J Dear, Franziska Kundel, Seema Qamar, Georg Meisl, Tuomas PJ Knowles, David Klenerman
Sep 20, 2018
aggregation mechanism amyloid oligomers Kinetic modeling single-molecule FRET Tau Amyloid beta-Peptides Codon Fluorescence Resonance Energy Transfer Genes Synthetic Humans Kinetics Models Molecular Neurofibrillary Tangles Polymers protein aggregation Pathological single molecule imaging tau Proteins
A global sampler of single particle tracking solutions for single molecule microscopy.
Michael Hirsch, Richard Wareham, Ji W Yoon, Daniel J Rolfe, Laura C Zanetti-Domingues, Michael P Hobson, Peter J Parker, Marisa L Martin-Fernandez, Sumeetpal S Singh
Sep 09, 2019
The dependence on model-fitting to evaluate particle trajectories makes it difficult for single particle tracking (SPT) to resolve the heterogeneous molecular motions typical of cells. We present here a global spatiotemporal...
Missed cleavage opportunities by FEN1 lead to Okazaki fragment maturation via the long-flap pathway.
Manal S Zaher, Fahad Rashid, Bo Song, Luay I Joudeh, Mohamed A Sobhy, Muhammad Tehseen, Manju M Hingorani, Samir M Hamdan
Sep 11, 2018
RNA-DNA hybrid primers synthesized by low fidelity DNA polymerase α to initiate eukaryotic lagging strand synthesis must be removed efficiently during Okazaki fragment (OF) maturation to complete DNA replication. In this...
Acetyltransferases DNA DNA Cleavage DNA Helicases DNA Replication DNA Fungal Fluorescence Resonance Energy Transfer Kinetics Membrane Proteins Protein Binding Replication Protein A Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Signal Transduction single molecule imaging Substrate Specificity
Graphene Liquid Enclosure for Single-Molecule Analysis of Membrane Proteins in Whole Cells Using Electron Microscopy.
Indra N Dahmke, Andreas Verch, Justus Hermannsdörfer, Diana B Peckys, Robert S Weatherup, Stephan Hofmann, Niels de Jonge
May 04, 2018
Membrane proteins govern many important functions in cells via dynamic oligomerization into active complexes. However, analytical methods to study their distribution and functional state in relation to the cellular structure are...
STEM breast cancer cell epidermal growth factor receptor graphene liquid-phase electron microscopy single-molecule analysis tunneling nanotube Cell Line Tumor Graphite Humans Lab-On-A-Chip Devices Membrane Proteins Microscopy Electron Neoplasm Proteins Quantum Dots Receptor ErbB-2 Silicon Compounds single molecule imaging
FRET-enhanced photostability allows improved single-molecule tracking of proteins and protein complexes in live mammalian cells.
Srinjan Basu, Lisa-Maria Needham, David Lando, Edward JR Taylor, Kai J Wohlfahrt, Devina Shah, Wayne Boucher, Yi Lei Tan, Lawrence E Bates, Olga Tkachenko, Julie Cramard, B Christoffer Lagerholm, Christian Eggeling, Brian Hendrich, Dave Klenerman, Steven F Lee, Ernest D Laue
Aug 14, 2018
A major challenge in single-molecule imaging is tracking the dynamics of proteins or complexes for long periods of time in the dense environments found in living cells. Here, we introduce the concept of using FRET to enhance the...
Superresolving the kidney-a practical comparison of fluorescence nanoscopy of the glomerular filtration barrier.
Lucia Cs Wunderlich, Florian Ströhl, Stefan Ströhl, Oliver Vanderpoorten, Luca Mascheroni, Clemens F Kaminski
Feb 12, 2021
Immunofluorescence microscopy is routinely used in the diagnosis of and research on renal impairments. However, this highly specific technique is restricted in its maximum resolution to about 250 nm in the lateral and 700 nm in...
Fluorescence microscopy formalin-fixed paraffin-embedded Kidney Nephrin Podocyte foot processes super-resolution microscopy Animals Glomerular Filtration Barrier Humans Kidney Membrane Proteins Mice Inbred C57BL Microscopy Fluorescence Paraffin Embedding Podocytes single molecule imaging Tissue Fixation
vLUME
Alexander Spark, Alexandre Kitching, Daniel Esteban-Ferrer, Anoushka Handa, Alexander R Carr, Lisa-Maria Needham, Aleks Ponjavic, Ana Mafalda Santos, James McColl, Christophe Leterrier, Simon J Davis, Ricardo Henriques, Steven F Lee
Sep 03, 2020
Atomic force microscopy for single molecule characterisation of protein aggregation.
The development of atomic force microscopy (AFM) has opened up a wide range of novel opportunities in nanoscience and new modalities of observation in complex biological systems. AFM imaging has been widely employed to resolve...
resPAINT
Edward W Sanders, Alexander R Carr, Ezra Bruggeman, Markus Körbel, Sarah I Benaissa, Robert F Donat, Ana M Santos, James McColl, Kevin O'Holleran, David Klenerman, Simon J Davis, Steven F Lee, Aleks Ponjavic
Aug 23, 2022
Points for accumulation in nanoscale topography (PAINT) allows practically unlimited measurements in localisation microscopy but is limited by background fluorescence at high probe concentrations, especially in volumetric...
Infrared nanospectroscopy reveals the molecular interaction fingerprint of an aggregation inhibitor with single Aβ42 oligomers.
Francesco Simone Ruggeri, Johnny Habchi, Sean Chia, Robert I Horne, Michele Vendruscolo, Tuomas PJ Knowles
Dec 09, 2020
Significant efforts have been devoted in the last twenty years to developing compounds that can interfere with the aggregation pathways of proteins related to misfolding disorders, including Alzheimer's and Parkinson's diseases....
Alzheimer Disease Amyloid beta-Peptides Bexarotene Drug Discovery Feasibility Studies Humans Hydrogen Bonding Kinetics Microscopy Atomic Force Peptide Fragments Protein Aggregates protein aggregation Pathological Recombinant Proteins single molecule imaging Spectrophotometry Infrared Structure-Activity Relationship Vibration
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