A5003690673- Advanced Electron Microscopy Techniques and Applications
- Advanced X-ray Imaging Techniques
- Liver physiology and pathology
- Electron and X-Ray Spectroscopy Techniques
- Advanced Fluorescence Microscopy Techniques
- Photoacoustic and Ultrasonic Imaging
- Nanoplatforms for cancer theranostics
- RNA and protein synthesis mechanisms
- Wnt/β-catenin signaling in development and cancer
- Caveolin-1 and cellular processes
- Metal-Organic Frameworks: Synthesis and Applications
- X-ray Spectroscopy and Fluorescence Analysis
- Cell Adhesion Molecules Research
- RNA modifications and cancer
- Enzyme Structure and Function
- Advanced MRI Techniques and Applications
- Bacteriophages and microbial interactions
- Herpesvirus Infections and Treatments
- Graphene and Nanomaterials Applications
- Vector-borne infectious diseases
- Advanced X-ray and CT Imaging
- RNA Interference and Gene Delivery
- Cancer, Hypoxia, and Metabolism
- Immune cells in cancer
- Endoplasmic Reticulum Stress and Disease
Centro Nacional de Biotecnología
2012-2024
ALBA Synchrotron (Spain)
2017-2023
Consejo Superior de Investigaciones Científicas
2015-2023
Max Planck Institute of Molecular Cell Biology and Genetics
2023
Queen Mary University of London
2023
William Harvey Research Institute
2023
Universidad Autónoma de Madrid
2022
Universidad Complutense de Madrid
2011
The influenza viruses cause annual epidemics of respiratory disease and occasional pandemics, which constitute a major public-health issue. segmented negative-stranded RNAs are associated with the polymerase complex nucleoprotein (NP), forming ribonucleoproteins (RNPs), responsible for virus transcription replication. We describe structure native RNPs derived from virions. They show double-helical conformation in two NP strands opposite polarity each other along helix. Both connected by...
Abstract Recent work in biomolecule‐metal–organic framework (MOF) composites has proven to be an effective strategy for the protection of proteins. However, other biomacromolecules such as nucleic acids, encapsulation into nano MOFs and related characterizations are their infancy. Herein, a complete gene‐set zeolitic imidazolate framework‐8 (ZIF‐8) cellular expression gene delivered by MOF reported. Using green fluorescent protein (GFP) plasmid (plGFP) proof‐of‐concept genetic macromolecule,...
Abstract Double-stranded DNA bacteriophages package their genome at high pressure inside a procapsid through the portal, an oligomeric ring protein located unique capsid vertex. Once has been packaged, tail components assemble on portal to render mature infective virion. The tightly seals ejection conduit until infection, when its interaction with host membrane triggers opening of channel and viral is delivered cell. Using high-resolution cryo-electron microscopy X-ray crystallography, here...
Abstract The iridium half‐sandwich complex [Ir(η 5 :κ 1 ‐C Me 4 CH 2 py)(2‐phenylpyridine)]PF 6 is highly cytotoxic: 15–250× more potent than clinically used cisplatin in several cancer cell lines. We have developed a correlative 3D cryo X‐ray imaging approach to specifically localize and quantify within the whole hydrated at nanometer resolution. By means of soft tomography (cryo‐SXT), which provides cellular ultrastructure 50 nm resolution, hard fluorescence (cryo‐XRF), elemental...
Recent advances in nanoparticle design have generated new possibilities for nano-biotechnology and nano-medicine. Here we used cryo-soft X-ray tomography (cryo-SXT) to collect comprehensive three-dimensional (3D) data characterise the interaction of superparamagnetic iron oxide nanoparticles (SPION) with a breast cancer cell line. We incubated MCF-7 (a human line) from 0 24 h SPION (15 nm average diameter, coated dimercaptosuccinic acid), system that has been studied previously using various...
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...
Cryo-electron microscopy (cryoEM) has become a well established technique to elucidate the 3D structures of biological macromolecules. Projection images from thousands macromolecules that are assumed be structurally identical combined into single map representing Coulomb potential macromolecule under study. This article discusses possible caveats along image-processing path and how avoid them obtain reliable structure. Some these problems very known in community. These may referred as...
The presence of preferred orientations in single particle analysis (SPA) by cryo-Electron Microscopy (cryoEM) is currently one the hurdles preventing many structural analyses from yielding high-resolution structures. Although existence mostly related to grid preparation, this technical note, we show that some image processing algorithms used for angular assignment and three-dimensional (3D) reconstruction are more robust than others these detrimental conditions. We exemplify argument with...
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.
Significance Platelet activation induces reorganization of the cell and formation cellular protrusions, pseudopodia. These processes are accompanied by remodeling actin cytoskeleton platelet integrins, which mediate strong adhesion to extracellular matrix. In this work, we analyzed polarity integrin architecture in A nonuniform filaments pseudopodia indicates that these protrusions may be involved contractile acto-myosin forces. Heterogeneity conformation was found, while solely a bent...
Viral cutaneous lesions are frequent in some bird populations, though we generally ignorant of the causal agent. In instances, they represent a threat to livestock and wildlife health. We present here multiplex PCR which detects distinguishes infection by two such agents, avipoxviruses papillomaviruses, avian hosts. assayed biopsies superficial skin swabs from field preserved museum specimens. Ninety-three percent samples symptomatic specimens tested positive for presence avipox (n = 23) or...
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.
Advances in cryo-electron microscopy (cryo-EM) have made it possible to obtain structures of large biological macromolecules at near-atomic resolution. This "resolution revolution" has encouraged the use and development modeling tools able produce high-quality atomic models from cryo-EM density maps. Unfortunately, many practical problems appear when combining different packages same processing workflow, which make difficult these by non-experts and, therefore, reduce their utility. We...
We report the synthesis of plasmonic nanocapsules and cellular responses they induce in 3D melanoma models for their perspective use as a photothermal therapeutic agent. The wall is composed polyelectrolytes. inner part functionalized with discrete gold nanoislands. cavity contains fluorescent payload to show ability loading cargo. exhibit simultaneous two-photon luminescent, properties X-ray contrasting ability. average fluorescence lifetime (τ) measured FLIM (0.3 ns) maintained regardless...
Biomineralization is the process by which living organisms generate organized mineral crystals. In human cells, this phenomenon culminates with formation of hydroxyapatite, a naturally occurring form calcium apatite. The mechanism that explains genesis within cell and propagation in extracellular matrix still remains largely unexplained, its characterization highly controversial, especially humans. fact, up to now, biomineralization core knowledge has been provided investigations on advanced...
Revealing the intracellular location of novel therapeutic agents is paramount for understanding their effect at cell ultrastructure level. Here, we apply a correlative cryo 3D imaging approach to determine fate designed protein-nanomaterial hybrid with antifibrotic properties that shows great promise in mitigating myocardial fibrosis. Cryo structured illumination microscopy (cryo-3D-SIM) pinpoints and soft X-ray tomography (cryo-SXT) reveals ultrastructural environment subcellular...
ABSTRACT During host cell invasion, Shigella escapes to the cytosol and polymerizes actin for cell-to-cell spread. To restrict spread, cells employ cell-autonomous immune responses including antibacterial autophagy septin cage entrapment. How septins interact with process target destruction is poorly understood. Here, we employed a correlative light cryo-soft X-ray tomography (cryo-SXT) pipeline study entrapment in its near-native state. Quantitative cryo-SXT showed that fragments...
Babesia is an apicomplexan parasite of significance that causes the disease known as babesiosis in domestic and wild animals humans worldwide. infects vertebrate hosts reproduces asexually by a form binary fission within erythrocytes/red blood cells (RBCs), yielding complex pleomorphic population intraerythrocytic parasites. Seven them, clearly visible human RBCs infected with divergens, are considered main forms named single, double, quadruple trophozoites, paired double pyriforms, tetrad...
Abstract Multimodal imaging promises to revolutionize the understanding of biological processes across scales in space and time by combining strengths multiple techniques. Fluorescent nanodiamonds (FNDs) are biocompatible, chemically inert, provide high contrast light‐ electron‐based microscopy, versatile optical quantum sensors. Here it is demonstrated that FNDs also absorption nanoscale 3D soft X‐ray tomograms with a resolution 28 nm all dimensions. Confocal fluorescence, atomic force,...
Macrophages are a heterogeneous population of innate immune cells that support tissue homeostasis through their involvement in development and repair, pathogen defense. Emerging data reveal metabolism may control macrophage polarization function and, conversely, phenotypic drive metabolic reprogramming.Here we use biochemical analysis, correlative cryogenic fluorescence microscopy cryo-focused ion-beam scanning electron microscopy.We demonstrate growth hormone (GH) reprograms inflammatory...
Abstract The iridium half‐sandwich complex [Ir(η 5 :κ 1 ‐C Me 4 CH 2 py)(2‐phenylpyridine)]PF 6 is highly cytotoxic: 15–250× more potent than clinically used cisplatin in several cancer cell lines. We have developed a correlative 3D cryo X‐ray imaging approach to specifically localize and quantify within the whole hydrated at nanometer resolution. By means of soft tomography (cryo‐SXT), which provides cellular ultrastructure 50 nm resolution, hard fluorescence (cryo‐XRF), elemental...
In the HIV vaccine field, there is a need to produce highly immunogenic forms of Env protein with capacity trigger broad B and T-cell responses. Here, we report generation characterization chimeric HIV-1 gp120 (termed gp120-14K) by fusing from clade vaccinia virus (VACV) 14K oligomeric (derived A27L gene). Stable CHO cell lines expressing gp120-14K were generated purified was characterized size exclusion chromatography, electron microscopy binding anti-Env antibodies. These approaches...
Epithelial intercellular adhesion molecule (ICAM)-1 is apically polarized, interacts with, and guides leukocytes across epithelial barriers. Polarized hepatic epithelia organize their apical membrane domain into bile canaliculi ducts, which are not accessible to circulating immune cells but that nevertheless confine most of ICAM-1. Here, by analyzing ICAM-1_KO human cells, liver organoids from mice rescue-of-function experiments, we show ICAM-1 regulates apicobasal polarity in a leukocyte...