Extracellular vesicle (EV) research increasingly demands for quantitative characterisation at the single vesicle level to address heterogeneity and complexity of EV subpopulations. Emerging, commercialised technologies for single EV analysis based on, for example, imaging flow cytometry or imaging after capture on chips generally require dedicated instrumentation and proprietary software not readily accessible to every lab. This limits their implementation for routine EV characterisation in the rapidly growing EV field. We and others have shown that single vesicles can be detected as light diffraction limited fluorescent spots using standard confocal and widefield fluorescence microscopes. Advancing this simple strategy into a process for routine EV quantitation, we developed 'EVAnalyzer', an ImageJ/Fiji (Fiji is just ImageJ) plugin for automated, quantitative single vesicle analysis from imaging data. Using EVAnalyzer, we established a robust protocol for capture, (immuno-)labelling and fluorescent imaging of EVs. To exemplify the application scope, the process was optimised and systematically tested for (i) quantification of EV subpopulations, (ii) validation of EV labelling reagents, (iii) in situ determination of antibody specificity, sensitivity and species cross-reactivity for EV markers and (iv) optimisation of genetic EV engineering. Additionally, we show that the process can be applied to synthetic nanoparticles, allowing to determine siRNA encapsulation efficiencies of lipid-based nanoparticles (LNPs) and protein loading of SiO2 nanoparticles. EVAnalyzer further provides a pipeline for automated quantification of cell uptake at the single cell-single vesicle level, thereby enabling high content EV cell uptake assays and plate-based screens. Notably, the entire procedure from sample preparation to the final data output is entirely based on standard reagents, materials, laboratory equipment and open access software. In summary, we show that EVAnalyzer enables rigorous characterisation of EVs with generally accessible tools. Since we further provide the plugin as open-source code, we expect EVAnalyzer to not only be a resource of immediate impact, but an open innovation platform for the EV and nanoparticle research communities.

• MeSH
• biologické markery metabolismus   MeSH
• diagnostické zobrazování MeSH
• extracelulární vezikuly * metabolismus   MeSH
• oxid křemičitý * metabolismus   MeSH
• průtoková cytometrie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• diagnostické zobrazování MeSH
• mamografie MeSH
• nemoci prsů MeSH
• prsy MeSH
• Publikační typ
• atlasy MeSH

• Konspekt
• Lékařské vědy. Lékařství
• NLK Obory
• endokrinologie
• radiologie, nukleární medicína a zobrazovací metody
• radiologie, nukleární medicína a zobrazovací metody

• MeSH
• fyzikální jevy MeSH
• magnetická rezonanční tomografie MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• radiologie, nukleární medicína a zobrazovací metody
• NLK Publikační typ
• kolektivní monografie

In the past decade, automated microscopy has become an important tool for the drug discovery and development process. The establishment of imaging modalities as screening tools depended on technological breakthroughs in the domain of automated microscopy and automated image analysis. These types of assays are often referred to as high content screening or high content analysis (HCS/HCA). The driving force to adopt imaging for drug development is the quantity and quality of cellular information that can be collected and the enhanced physiological relevance of cellular screening compared to biochemical screening. Most imaging in drug development is performed on fixed cells as this allows uncoupling the preparation of the cells from the acquisition of the images. Live-cell imaging is technically challenging, but is very useful for many aspects of the drug development pipeline such as kinetic studies of compound mode of action or to analyze the motion of cellular components. Most vendors of HCS microscopy systems offer the option of environmental chambers and onboard pipetting on their platforms. This reflects the wish and desire of many customers to have the ability to perform live-cell assays on their HCS automated microscopes. This book chapter summarizes the challenges and advantages of live-cell imaging in drug discovery. Examples of applications are presented and the motivation to perform these assays in kinetic mode is discussed.

• MeSH
• buněčné kultury MeSH
• kultivované buňky MeSH
• lidé MeSH
• mikroskopie MeSH
• molekulární zobrazování metody   MeSH
• objevování léků * metody   MeSH
• počítačové zpracování obrazu MeSH
• rychlé screeningové testy * MeSH
• software MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Zavedení a rozšíření elastografických metod zásadně změnilo klinickou hepatologickou praxi. Elastografické vyšetření je stále dostupnější a pole neinvazivní diagnostiky jaterních chorob se i nadále dynamicky rozvíjí. Objevují se nové technologie i indikace umožňující neinvazivní staging jaterních chorob v mnohem větším rozsahu. Oblasti elastografie jater v současnosti dominují metody založené na využití ultrazvuku – transientní elastografie (TE) a shear wave elastografie (SWE). Obě metody prokazatelně diagnostikují pokročilou jaterní fibrózu/cirhózu s vysokou spolehlivostí u nejčastějších chronických jaterních chorob. Předpokladem validního výsledku je technicky správně provedené vyšetření a jeho interpretace v klinickém kontextu. TE navíc umožňuje hodnocení portální hypertenze u pacientů s kompenzovanou jaterní cirhózou. Novými metodami na poli neinvazivní diagnostiky jaterních chorob jsou na ultrazvuku založená kvantifikace jaterní steatózy, elastografie sleziny k hodnocení portální hypertenze a elastografie ložisek v játrech.

Introduction and spread of elastographical methods have changed clinical practice in hepatology substantially. Elastography is becoming more available and the field of non-invasive diagnostics of liver diseases keeps growing dynamically. New technologies and applications are being developed allowing non-invasive staging of liver diseases to much broader extent. Ultrasound based methods like transient elastography (TE) and shear wave elastography (SWE) are dominating the field of liver elastography. Both methods are able to distinguish advanced liver fibrosis/ cirrhosis with high accuracy and in patients with all most common chronic liver diseases. A technically well performed examination and its interpretation in a clinical context are prerequisites for a valid diagnosis. TE also enables assessment of presence/severity of portal hypertension in patients with compensated chronic advanced liver disease. There are new methods under development and validation like an ultrasound-based liver fat content quantification, assessment of portal hypertension using elastography of the spleen or the use of elastography in the diagnostics of focal liver lesions.

• MeSH
• elastografie metody   přístrojové vybavení   MeSH
• jaterní cirhóza diagnostické zobrazování   MeSH
• lidé MeSH
• nemoci jater * diagnostické zobrazování   MeSH
• portální hypertenze MeSH
• ultrasonografie metody   MeSH
• ztučnělá játra diagnostické zobrazování   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

Magnetic resonance imaging (MRI) relies on appropriate contrast agents, especially for visualizing transplanted cells within host tissue. In recent years, compounds containing fluorine-19 have gained significant attention as MRI probe, particularly in dual 1H/19F-MR imaging. However, various factors affecting probe sensitivity, such as fluorine content and the equivalency of fluorine atoms, must be considered. In this study, we synthesized fluorinated micelles with adjustable surface positive charge density and investigated their physicochemical properties and MRI efficacy in phantoms and labeled cells. While the micelles exhibited clear signals in 19F-MR spectra and imaging, the concentrations required for MRI visualization of labeled cells were relatively high, adversely affecting cell viability. Despite their favourable physicochemical properties, achieving higher labeling rates without compromising cell viability during labeling remains a challenge for potential in vivo applications.

• MeSH
• barvení a značení metody   MeSH
• fantomy radiodiagnostické MeSH
• fluor chemie   MeSH
• halogenace MeSH
• kationty * chemie   MeSH
• kontrastní látky chemie   MeSH
• lidé MeSH
• magnetická rezonanční tomografie metody   MeSH
• micely * MeSH
• myši MeSH
• viabilita buněk * účinky léků   MeSH
• zobrazování fluorovou magnetickou rezonancí metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• chorobopisy - počítačové systémy MeSH
• diagnostické zobrazování MeSH
• management znalostí MeSH
• metody pro podporu rozhodování MeSH
• počítačové zpracování signálu MeSH
• řízení zdravotnictví MeSH
• studium lékařství MeSH
• výpočetní biologie MeSH
• zdravotnické informační systémy MeSH
• Publikační typ
• sborníky MeSH

• Konspekt
• Lékařské vědy. Lékařství
• NLK Obory
• lékařská informatika
• NLK Publikační typ
• ročenky

• MeSH
• diagnostické zobrazování MeSH
• radiografie MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• radiologie, nukleární medicína a zobrazovací metody
• NLK Publikační typ
• učebnice vysokých škol
• kolektivní monografie

Magnetic resonance imaging (MRI) enables a noninvasive in vivo quantification of iron in various organs. Several techniques have been developed that detect signal alterations derived mainly from the magnetic properties of ferritin and hemosiderin, the major iron storage compounds. High magnetic susceptibility of ferritin shortens the transversal relaxation time of nearby water protons and thus induces a focal signal extinction of iron-rich areas in T2-weighted (T2w) MRI. T2w tissue contrast is additionally influenced by other factors such as water content, myelin density, and the presence of other metals. Therefore, more specific methods are needed with higher specificity to iron. These in vivo techniques can be divided into three groups: relaxometry, magnetic field correlation imaging and phase-based contrast covering susceptibility-weighted imaging, and quantitative susceptibility mapping. The differential diagnosis of various neurological disorders is aided by characteristic patterns of iron depositions. Reliable estimates of cerebral tissue iron concentration are equally important in studying physiological age-related as well as pathological conditions in neurodegenerative, neuroinflammatory, and vascular diseases. In the future, monitoring changes in iron storage and content may serve as sensitive biomarker for diagnosis as well as treatment monitoring.

• MeSH
• degenerace nervu metabolismus   MeSH
• lidé MeSH
• magnetická rezonanční tomografie * MeSH
• mozek metabolismus   MeSH
• neurodegenerativní nemoci metabolismus   MeSH
• železo metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Over the last few years, the development and relevance of 19F magnetic resonance imaging (MRI) for use in clinical practice has emerged. MRI using fluorinated probes enables the achievement of a specific signal with high contrast in MRI images. However, to ensure sufficient sensitivity of 19F MRI, fluorine probes with a high content of chemically equivalent fluorine atoms are required. The majority of 19F MRI agents are perfluorocarbon emulsions, which have a broad range of applications in molecular imaging, although the content of fluorine atoms in these molecules is limited. In this review, we focus mainly on polymer probes that allow higher fluorine content and represent versatile platforms with properties tailorable to a plethora of biomedical in vivo applications. We discuss the chemical development, up to the first imaging applications, of these promising fluorine probes, including injectable polymers that form depots that are intended for possible use in cancer therapy.

• MeSH
• fluor chemie   MeSH
• fluorokarbony chemie   MeSH
• koncentrace vodíkových iontů MeSH
• kontrastní látky chemie   MeSH
• lidé MeSH
• molekulární sondy chemie   MeSH
• molekulární zobrazování přístrojové vybavení   metody   MeSH
• myši MeSH
• polymery chemie   MeSH
• radiační rozptyl MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• světlo MeSH
• teplota MeSH
• zobrazování fluorovou magnetickou rezonancí metody   trendy   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH