A5046989784- Lipid Membrane Structure and Behavior
- Advanced Fluorescence Microscopy Techniques
- Spectroscopy and Quantum Chemical Studies
- Air Quality and Health Impacts
- RNA Interference and Gene Delivery
- Nanoparticles: synthesis and applications
- Nanopore and Nanochannel Transport Studies
- Spectroscopy Techniques in Biomedical and Chemical Research
- Force Microscopy Techniques and Applications
- Electron Spin Resonance Studies
- Inhalation and Respiratory Drug Delivery
- Optical Imaging and Spectroscopy Techniques
- HIV Research and Treatment
- Peroxisome Proliferator-Activated Receptors
- Luminescence and Fluorescent Materials
- Analytical Chemistry and Sensors
- Photosynthetic Processes and Mechanisms
- Immune Response and Inflammation
- RNA Research and Splicing
- Protein Structure and Dynamics
- Nanoplatforms for cancer theranostics
- Luminescence Properties of Advanced Materials
- 3D Printing in Biomedical Research
- Immune cells in cancer
- nanoparticles nucleation surface interactions
Jožef Stefan Institute
2016-2025
University of Oxford
2017-2024
MRC Weatherall Institute of Molecular Medicine
2018-2024
MRC Human Immunology Unit
2017-2022
Institute of Molecular Medicine
2020
Science Oxford
2020
John Radcliffe Hospital
2018
University of Ljubljana
2008
Abstract Regulation of plasma membrane curvature and composition governs essential cellular processes. The material property bending rigidity describes the energetic cost deformations depends on molecular composition. Because compositional fluctuations active processes, it is challenging to measure in intact cells. Here, we study using giant vesicles (GPMVs), which largely preserve lipidome proteome. We show that membranes under varied conditions correlated readout from environment-sensitive...
The structural diversity of different lipid species within the membrane defines its biophysical properties such as fluidity, phase transition, curvature, charge distribution, and tension. Environment-sensitive probes, which change their spectral in response to surrounding milieu, have greatly contributed our understanding properties. To realize full potential these probes avoid misinterpretation responses, a detailed investigation fluorescence characteristics environments is necessary. Here,...
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...
Although the link between inhalation of nanoparticles and cardiovascular disease is well established, causal pathway nanoparticle exposure increased activity blood coagulation factors remains unexplained. To initiate tissue factor bearing epithelial cell membranes should be exposed to blood, on other side less than a micrometre thin air-blood barrier. For inhaled promote coagulation, they need bind lung epithelial-cell membrane parts relocate them into blood. assess this hypothesis, we use...
To disentangle the elusive lipid–protein interactions in T‐cell activation, we investigate how externally imposed variations mobility of key membrane proteins (T‐cell receptor [TCR], kinase Lck, and phosphatase CD45) affect local lipid order protein colocalisation. Using spectral imaging with polarity‐sensitive probes model membranes live Jurkat T cells, find that partial immobilisation (including TCR) by aggregation or ligand binding changes their preference towards a more ordered...
It is important to understand the nanomaterials intracellular trafficking and distribution investigate their targeting into nuclear area in living cells. In our previous study, we firstly observed penetration of nonmodified positively charged carbon dots decorated with quaternary ammonium groups (QCDs) nucleus mouse NIH/3T3 fibroblasts. Thus, this work, focused on deeper study QCDs inside two healthy L929 cell lines by fluorescence microspectroscopy performed a comprehensive cytotoxic DNA...
Abstract On a daily basis, people are exposed to multitude of health‐hazardous airborne particulate matter with notable deposition in the fragile alveolar region lungs. Hence, there is great need for identification and prediction material‐associated diseases, currently hindered due lack in‐depth understanding causal relationships, particular between acute exposures chronic symptoms. By applying advanced microscopies omics vitro vivo systems, together silico molecular modeling, it determined...
Molecular plasma membrane organization and dynamics play an important role in cellular signalling. Advances our understanding of the nanoscale architecture heavily rely on development non-invasive experimental methods, particularly advanced fluorescence microscopy spectroscopy techniques with high spatio-temporal resolution sensitivity to local molecular properties. However, it remains difficult combine several them for a multimodal characterisation that would reduce possibility...
Fluorescence correlation spectroscopy in combination with super-resolution stimulated emission depletion microscopy (STED-FCS) is a powerful tool to investigate molecular diffusion sub-diffraction resolution.It has been of particular use for investigations two dimensional systems like cell membranes, but so far seen very limited applications studies three-dimensional diffusion.One reason this the extreme sensitivity axial (z) STED pattern optical aberrations.We present here an adaptive...
Super-resolution stimulated emission depletion (STED) microcopy provides optical resolution beyond the diffraction limit. The can be increased laterally (xy) or axially (z). Two-dimensional STED has been extensively used to elucidate nanoscale membrane structure and dynamics via imaging combined with spectroscopy techniques such as fluorescence correlation (FCS) spectral imaging. On contrary, z-STED not in this context. Here, we show that a combination of FCS enables us see previously...
Abstract Fluorescence microscopy is limited by photoconversion due to continuous illumination, which results in not only photobleaching but also conversion of fluorescent molecules into species different spectral properties through photoblueing. Here, we determined fluorescence parameters photoconverted products for various fluorophores under standard confocal and stimulated emission depletion (STED) conditions. We observed changes both spectra lifetimes that can cause artifacts quantitative...
Despite many advances in gene therapy, the delivery of small interfering RNAs is still challenging. Erythrocytes are most abundant cells human body, and their membrane possesses unique features. From them, erythrocytes vesicles can be generated, employable as nano drug system with prolonged blood residence high biocompatibility. Human erythrocyte ghosts were extruded presence siRNA, objects termed EMVs (erythrocyte vesicles). An ultracentrifugation-based method was applied to select only...
A photoswitchable quencher can be used to reversibly turn off the emission from a fluorescent dye, generating small molecule dyad that is effective for super-resolution RESOLFT microscopy.
Fluorescence microspectroscopy (FMS) with environmentally sensitive dyes provides information about local molecular surroundings at microscopic spatial resolution. Until recently, only probes exhibiting large spectral shifts due to changes have been used. For filter-based experimental systems, where signal different wavelengths is acquired sequentially, photostability has required in addition. Herein, we systematically analyzed our fitting models and bleaching correction algorithms which...
Lack of better understanding nanoparticles targeted delivery into cancer cells calls for advanced optical microscopy methodologies.Here we present a development fluorescence microspectroscopy (spectral imaging) based on white light spinning disk confocal microscope with emission wavelength selection by liquid crystal tunable filter.Spectral contrasting images was used to localize polymer and cell membranes labeled fluorophores that have substantially overlapping spectra.In addition, enabled...
Abstract Probing the diffusion of molecules has become a routine measurement across life sciences, chemistry and physics. It provides valuable insights into reaction dynamics, oligomerisation, molecular (re-)organisation or cellular heterogeneities. Fluorescence correlation spectroscopy (FCS) is one widely applied techniques to determine dynamics in two three dimensions. This technique relies on temporal autocorrelation intensity fluctuations but recording these thus far been limited by...
Several biopersistent high aspect ratio nanomaterials show pronounced pathogenic effects, from chronic lung inflammation and fibrosis to cancer. For example, asbestos fibers are classified as carcinogens, whereas the carcinogenicity of highly inflammatory multi-wall carbon nanotubes (MWCNTs), TiO2 is still being evaluated. The exact early mechanisms their pathogenicity towards cancer remains uncertain, but it likely not due genotoxic or mutagenic activity. A proposed mode action that might...
Cellular functions rely on proper actions of organelles such as peroxisomes. These the import proteins from cytosol. The peroxisomal receptor PEX5 takes up target in cytosol and transports them to matrix. However, its cytosolic molecular interactions have so far not directly been disclosed. Here, we combined advanced optical microscopy spectroscopy techniques fluorescence correlation stimulated emission depletion with biochemical tools present a detailed characterization diffusion...