A5000081681- Advanced Electron Microscopy Techniques and Applications
- Electron and X-Ray Spectroscopy Techniques
- Advanced X-ray Imaging Techniques
- Advanced Optical Imaging Technologies
- Optical Polarization and Ellipsometry
- Image Processing Techniques and Applications
- X-ray Spectroscopy and Fluorescence Analysis
- Enzyme Structure and Function
- Advanced optical system design
- Optical Coherence Tomography Applications
- Advanced Fluorescence Microscopy Techniques
- SARS-CoV-2 and COVID-19 Research
- Advanced MRI Techniques and Applications
- Medical Imaging Techniques and Applications
- Visual perception and processing mechanisms
- Force Microscopy Techniques and Applications
- RNA modifications and cancer
- Machine Learning in Materials Science
- Advanced X-ray and CT Imaging
- Liquid Crystal Research Advancements
- Cell Image Analysis Techniques
- Endoplasmic Reticulum Stress and Disease
- Lipid Membrane Structure and Behavior
- Integrated Circuits and Semiconductor Failure Analysis
- Heat shock proteins research
MRC Laboratory of Molecular Biology
2019-2023
ALBA Synchrotron (Spain)
2022-2023
Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas
2023
Max Planck Institute of Biochemistry
2023
Medical Research Council
2020
Centro Nacional de Biotecnología
2012-2019
Universidad Autónoma de Madrid
2012-2016
Biocom
2016
Consejo Superior de Investigaciones Científicas
2012-2015
Tel Aviv University
2013
We present a new approach for macromolecular structure determination from multiple particles in electron cryo-tomography (cryo-ET) data sets. Whereas existing subtomogram averaging approaches are based on 3D models, we propose to optimise regularised likelihood target that approximates function of the 2D experimental images. In addition, analogous Bayesian polishing and contrast transfer (CTF) refinement single-particle analysis, describe exploit increased signal-to-noise ratio averaged...
Abstract Motivation: Structural information of macromolecular complexes provides key insights into the way they carry out their biological functions. Achieving high-resolution structural details with electron microscopy requires identification a large number (up to hundreds thousands) single particles from micrographs, which is laborious task if it has be manually done and constitutes hurdle towards high-throughput. Automatic particle selection in micrographs far being settled new more...
Full field soft X-ray microscopy is becoming a powerful imaging technique to analyze whole cells preserved under cryo conditions. Images obtained in these microscopes can be combined by tomographic reconstruction quantitatively estimate the three-dimensional (3D) distribution of absorption coefficients inside cell. The impulse response an system one factors that limits quality microscope reconstructions. main goal this work experimentally measure 3D and assess optical resolution depth...
Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virions are surrounded by a lipid bilayer from which spike (S) protein trimers protrude. Heavily glycosylated S bind the ACE2 receptor and mediate entry of into target cells. exhibits extensive conformational flexibility: it modulates exposure its binding site later undergoes complete structural rearrangement to drive fusion viral cellular membranes. The structures conformations soluble, overexpressed, purified proteins...
Image processing in cryogenic electron tomography (cryoET) is currently at a similar state as Single Particle Analysis (SPA) microscopy (cryoEM) was few years ago. Its data workflows are far from being well defined and the user experience still not smooth. Moreover, file formats of different software packages their associated metadata standardized, mainly since developed by groups, focusing on steps pipeline. The Scipion framework, originally for SPA (de la Rosa-Trevín et al., 2016), has...
The inherent distortion of a reflective parallel aligned spatial light modulator (SLM) may need compensation not only for the backplane curvature but also other possible nonuniformities caused by thickness variations liquid crystal layer across aperture. First, we build global look-up table (LUT) phase modulation versus addressed gray level whole device Second, when lack uniformity is observed, define grid cells onto SLM aperture and develop multipoint calibration. relative between...
Abstract We present a new approach for macromolecular structure determination from multiple particles in electron cryo-tomography (cryo-ET) data sets. Whereas existing subtomogram averaging approaches are based on 3D models, we propose to optimise regularised likelihood target that approximates function of the 2D experimental images. In addition, analogous Bayesian polishing and contrast transfer (CTF) refinement single-particle analysis, describe exploit increased signal-to-noise ratio...
New image processing methodologies and algorithms have greatly contributed to the signi cant progress in three-dimensional electron microscopy (3DEM) of biological complexes we seen over last decades. Naturally, availability accurate procedures for objective testing new is a crucial requirement further advancement eld. A good accepted work ow involves generation realistic 3DEM-like maps macromolecules from which some measure ground truth can be derived, ideally because their 3D atomic...
We used soft X-ray three-dimensional imaging to quantify the mass of superparamagnetic iron oxide nanoparticles (SPION) within whole cells, by exploiting differential absorption contrast. Near-edge nanotomography (NEASXT) combines whole-cell 3D structure determination at 50 nm resolution, with elemental mapping and high throughput. detected distribution SPIONs cells 0.3 g/cm(3) sensitivity, sufficient for detecting density corresponding a single nanoparticle.
Eukaryotic chaperonin-containing TCP1 drives tubulin dynamics and centrosome structure at immune synaptic contacts.
Abstract We have developed a new data collection method and processing framework in full field cryo soft X-ray tomography to computationally extend the depth of (DOF) Fresnel zone plate lens. Structural features 3D-reconstructed eukaryotic cells that are affected by DOF artifacts standard reconstruction now recovered. This approach, based on focal series projections, is easily applicable with closed expressions select specific acquisition parameters.
A proposal to dynamically compensate chromatic aberration of a programmable phase Fresnel lens displayed on liquid crystal device and working under broadband illumination is presented. It based time multiplexing set lenses, designed with common focal length for different wavelengths, tunable spectral filter that makes each sublens work almost monochromatically. Both the by spatial light modulator are synchronized. The whole sublenses within integration sensor. As result central order...
Soft X-ray tomography (SXT) is becoming a powerful imaging technique to analyze eukaryotic whole cells close their native state. Central the analysis of quality SXT 3D reconstruction estimation spatial resolution and Depth Field microscope. In turn, characterization Modulation Transfer Function (MTF) optical system key calculate both parameters. Consequently, in this work we introduce fully automated accurately estimate transfer function such an system. Our proposal based on preprocessing...
Abstract Electron Microscopy is reaching new capabilities thanks to the combined effect of technologies and image processing methods. However, reconstruction process still complex, requiring many steps elaborated optimization procedures. Therefore, possibility reach a wrong structure exists, justifying need robust statistical tests. In this work, we present conceptually simple alignment test, which does not require tilt-pair images, evaluate consistency between set projection images with...