The necessity to scrutinize more and biological molecules interactions both in solution on the cellular level has led an increasing demand for sensitive specific multiplexed diagnostic analysis. Photoluminescence (PL) detection is ideally suited biosensing bioimaging because it rapid there almost unlimited choice of fluorophores that provide a large versatility photophysical properties, including PL intensities, spectra, lifetimes.The most frequently used technique detect multiple parameters...

Fluorescence barcoding based on nanoparticles provides many advantages for multiparameter imaging. However, creating different concentration-independent codes without mixing various and by using single-wavelength excitation emission multiplexed cellular imaging is extremely challenging. Herein, we report the development of quantum dots (QDs) with two SiO

The time-gated FRET technique is used for rapid, sensitive intra- and extracellular imaging.

Imaging single molecules and nanoparticles in complex biological media is highly challenging notably due to autofluorescence of cells tissues. Lanthanides lanthanide complexes, with their particularly long luminescence lifetimes, offer the possibility perform time-gated imaging thus strongly reduce background. However, very low brightness photon flux have limited use single-molecule imaging. Here, we encapsulate high amounts Europium complexes into poly(methyl methacrylate)-based particles...

ABSTRACT Focal adhesions are composed of transmembrane integrins, linking the extracellular matrix to actomyosin cytoskeleton, via cytoplasmic proteins. Adhesion depends on activation integrins. Talin and kindlin proteins intracellular activators integrins that bind β-integrin tails. Integrin clustering through ligands guide organization adhesion complexes. However, roles talin in this process poorly understood. To determine contribution talin, kindlin, lipids integrin clustering, we used a...

A Atenção Primária à Saúde, segundo a Organização Mundial da Saúde é um cuidado essencial, baseado em métodos acessíveis, cientificamente validados e sustentáveis para comunidade, promovendo autoconfiança autodeterminação. O objetivo desse estudo foi analisar discrepância entre os processos programados efetivamente realizados duas Unidades Básicas de localizadas no município do Teresópolis/RJ. Estudo realizado período agosto novembro 2024 que revelou dificuldades significativas na...

Pulmonary hypertension (PH) is characterized by a progressive elevation in mean pulmonary arterial pressure, often leading to right ventricular failure and death. Growth factors play significant roles the pathogenesis of PH, their targeting may therefore offer novel therapeutic strategies this disease.To evaluate nerve growth factor (NGF) as potential new target PH.Expression and/or activation NGF its receptors were evaluated rat experimental PH induced chronic hypoxia or monocrotaline human...

Abstract The zebrafish is an important vertebrate model for disease, drug discovery, toxicity, embryogenesis, and neuroscience. In vivo fluorescence microscopy can reveal cellular subcellular details down to the molecular level with fluorescent proteins (FPs) currently main tool imaging. However, long maturation times, low brightness, photobleaching, broad emission spectra, sample autofluorescence are disadvantages that cannot be easily overcome by FPs. Here, a bright photostable...

Fluorescence signal enhancement via isothermal nucleic acid amplification is an important approach for sensitive imaging of intra- or extracellular protein biomarkers. Rolling circle (RCA) frequently applied fluorescence in situ but faces limitations concerning multiplexing, dynamic range, and the required multiple washing steps before imaging. Here, we show that Förster resonance energy transfer (FRET) between fluorescent dyes lanthanide (Ln) complexes hybridize to β-actin-specific RCA...

Fluorescent nanoparticles (NPs) have become irreplaceable tools for advanced cellular and subcellular imaging. While very bright NPs require excitation with UV or visible light, which can create strong autofluorescence of biological components, NIR-excitable without issues exhibit much lower brightness. Here, we show the application a new type surface-photosensitized terbium (Tb-NPs) autofluorescence-free intracellular imaging in live HeLa cells. The combination exceptionally high...

Abstract Lanthanide‐doped nanoparticles (LnNPs) have become an important class of fluorophores for advanced biosensing and bioimaging. LnNPs that are photosensitized by surface‐attached antenna ligands can possess exceptional brightness. However, their functional bioconjugation remains challenge translation into bioanalytical applications. To solve this problem, we designed a ligand be simultaneously applied as efficient light harvesting Tb surface ions strong linker biomolecules to the...

We present a simple modification of standard total internal reflection fluorescence microscope to achieve nanometric axial resolution, typically ≈10 nm. The technique is based on normalization images by conventional epi-illumination images. demonstrate the potential our method study adhesion phopholipid giant unilamellar vesicles.

Understanding the mechanisms of assembly and disassembly macromolecular structures in cells relies on solving biomolecular interacFons. However, those interacFons o_en remain unclear because tools to track molecular dynamics are not sufficiently resolved Fme or space. In this study, we present a straighaorward method for resolving inter- intra- adhesive machinery, using Quantum Dot based Förster Resonance Energy Transfer (FRET) nanosensors. IniFally, measured interacFon between Talin,...

Understanding the mechanisms of assembly and disassembly macromolecular structures in cells relies on solving biomolecular interactions. However, those interactions often remain unclear because tools to track molecular dynamics are not sufficiently resolved time or space. In this study, we present a straightforward method for resolving inter- intra- cell adhesive machinery, using quantum dot (QD) based Förster resonance energy transfer (FRET) nanosensors. Using mechanosensitive protein...

Abstract Understanding the mechanisms of assembly and disassembly macromolecular structures in cells relies on solving biomolecular interactions. However, those interactions often remain unclear because tools to track molecular dynamics are not sufficiently resolved time or space. In this study, we present a straightforward method for resolving inter‐ intra‐molecular cell adhesive machinery, using quantum dot (QD) based Förster resonance energy transfer (FRET) nanosensors. Using...

Abstract Fluoreszenzstrichkodierung mittels Nanopartikeln bietet viele Vorteile für die Multiparameter‐Mikroskopie. Es ist jedoch extrem schwierig, verschiedene konzentrationsunabhängige Codes zu erstellen, ohne Nanopartikel mischen und Anregung Emission einer einzelnen Wellenlänge zelluläre Multiplex‐Bildgebung verwenden. Hier berichten wir über Entwicklung von Quantenpunkten (quantum dots: QDs) mit zwei unterschiedlichen SiO 2 ‐Schalenstärken (6 12 nm), verschiedenen Lanthanoidkomplexen...

We propose a calibration routine useful to evaluate the incident angle in total internal reflection fluorescence (TIRF) microscopy. This procedure is based on critical measurements conducted back focal plane (BFP) of objective. Such BFP imaging can be easily implemented any TIRF setup, making this technique very attractive. Calibration exactitude was demonstrated by comparing theoretical angular dependence electric field intensity |E|2 at glass/water interface experimental observations.

Förster resonance energy transfer (FRET) between fluorescent dyes is a frequently applied technique for analyzing concentrations and conformations of biomolecules. Optimizing FRET by controlled dye-surface functionalization an important requirement to develop sensors based on surface–biomolecule interactions. Here, we investigate the silanization silica with aminosilanes ((3-aminopropyl)triethoxysilane, APTES) their subsequent commercial organic fluorophores (ATTO-550 ATTO-647N) controlling...

We propose a new strategy to evaluate adhesion strength at the single cell level. This approach involves variable-angle total internal reflection fluorescence microscopy monitor in real time topography of membranes, i.e. map membrane/substrate separation distance. According <em>Boltzmann</em> distribution, both potential energy profile and dissociation related interactions between membrane substrate were determined from topography. have highlighted on glass substrates coated with...

Abstract Understanding the mechanisms of assembly and disassembly macromolecular structures in cells relies on solving biomolecular interactions. However, those interactions often remain unclear because tools to track molecular dynamics are not sufficiently resolved time or space. In this study, we present a straightforward method for resolving inter‐ intra‐molecular cell adhesive machinery, using quantum dot (QD) based Förster resonance energy transfer (FRET) nanosensors. Using...