Super-resolution (SR) microscopy is a cutting-edge method that can provide detailed structural information with high resolution. However, the thickness of the specimen has been a major limitation for SR methods, and large biological structures have posed a challenge. To overcome this, the key step is to optimise sample preparation to ensure optical homogeneity and clarity, which can enhance the capabilities of SR methods for the acquisition of thicker structures. Oocytes are the largest cells in the mammalian body and are crucial objects in reproductive biology. They are especially useful for studying membrane proteins. However, oocytes are extremely fragile and sensitive to mechanical manipulation and osmotic shocks, making sample preparation a critical and challenging step. We present an innovative, simple and sensitive approach to oocyte sample preparation for 3D STED acquisition. This involves alcohol dehydration and mounting into a high refractive index medium. This extended preparation procedure allowed us to successfully obtain a unique two-channel 3D STED SR image of an entire mouse oocyte. By optimising sample preparation, it is possible to overcome current limitations of SR methods and obtain high-resolution images of large biological structures, such as oocytes, in order to study fundamental biological processes. Lay Abstract: Super-resolution (SR) microscopy is a cutting-edge tool that allows scientists to view incredibly fine details in biological samples. However, it struggles with larger, thicker specimens, as they need to be optically clear and uniform for the best imaging results. In this study, we refined the sample preparation process to make it more suitable for SR microscopy. Our method includes carefully dehydrating biological samples with alcohol and then transferring them into a mounting medium that enhances optical clarity. This improved protocol enables high-resolution imaging of thick biological structures, which was previously challenging. By optimizing this preparation method, we hope to expand the use of SR microscopy for studying large biological samples, helping scientists better understand complex biological structures.

• MeSH
• alkoholy MeSH
• fluorescenční mikroskopie metody   MeSH
• myši MeSH
• odběr biologického vzorku metody   MeSH
• oocyty * MeSH
• refraktometrie metody   MeSH
• zobrazování trojrozměrné * metody   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

PURPOSE: Fuchs endothelial corneal dystrophy (FECD) is a common, age-related cause of visual impairment. This systematic review synthesizes evidence from the literature on artificial intelligence (AI) models developed for the diagnosis and management of FECD. METHODS: We conducted a systematic literature search in MEDLINE, PubMed, Web of Science, and Scopus from January 1, 2000, to June 31, 2024. Full-text studies utilizing AI for various clinical contexts of FECD management were included. Data extraction covered model development, predicted outcomes, validation, and model performance metrics. We graded the included studies using the Quality Assessment of Diagnostic Accuracies Studies 2 tool. This review adheres to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) recommendations. RESULTS: Nineteen studies were analyzed. Primary AI algorithms applied in FECD diagnosis and management included neural network architectures specialized for computer vision, utilized on confocal or specular microscopy images, or anterior segment optical coherence tomography images. AI was employed in diverse clinical contexts, such as assessing corneal endothelium and edema and predicting post-corneal transplantation graft detachment and survival. Despite many studies reporting promising model performance, a notable limitation was that only three studies performed external validation. Bias introduced by patient selection processes and experimental designs was evident in the included studies. CONCLUSIONS: Despite the potential of AI algorithms to enhance FECD diagnosis and prognostication, further work is required to evaluate their real-world applicability and clinical utility. TRANSLATIONAL RELEVANCE: This review offers critical insights for researchers, clinicians, and policymakers, aiding their understanding of existing AI research in FECD management and guiding future health service strategies.

• MeSH
• Fuchsova endoteliální dystrofie * diagnóza   terapie   MeSH
• lidé MeSH
• optická koherentní tomografie metody   MeSH
• umělá inteligence * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• systematický přehled MeSH

OBJECTIVES: This study aimed to investigate the impact of bleaching agents based on carbamide or hydrogen peroxide on dental ceramics in vitro, utilizing scanning electron microscopy (SEM) and elemental analysis via inductively coupled plasma optical emission spectroscopy (ICP-OES). METHODS: CAD/CAM ceramics (IPS e.max®CAD, IPS Empress®CAD, Vitablocs® Mark II, Celtra Duo, and inCoris TZI) were treated with bleaching agents using either 10%, 20%, 30% carbamide peroxide or with 35%, and 40% hydrogen peroxide. RESULTS: Surface elemental release was not significantly affected by the type or concentration of bleaching agent (p>0.05). Ion release in feldspathic ceramics was significantly higher than in other ceramic materials (p⟨0.0001). Microstructural surface changes were observed in all materials except for lithium disilicate and zirconia-reinforced lithium silicate ceramics. CONCLUSIONS: All bleaching agents tested in this study showed a similar impact within each material type tested regarding total mass loss, elemental composition, or surface structure. CLINICAL RELEVANCE: Lithium disilicate and zirconia-reinforced lithium silicate ceramics were the most resistant to bleaching agents. In contrast, feldspathic ceramic showed the highest ion release and surface deterioration when exposed to all bleaching agents tested.

• MeSH
• design s pomocí počítače * MeSH
• karbamidperoxid * chemie   MeSH
• keramika * chemie   MeSH
• látky na bělení zubů * chemie   MeSH
• mikroskopie elektronová rastrovací MeSH
• peroxid vodíku * chemie   MeSH
• povrchové vlastnosti MeSH
• testování materiálů MeSH
• zirkonium chemie   MeSH
• zubní porcelán * chemie   MeSH
• Publikační typ
• časopisecké články MeSH

Atlas, který se zaměřuje na anatomii lidského mozku, zobrazenou mikroskopicky, makroskopicky a pomocí magnetické rezonance. Určeno odborné veřejnosti.; Učebnice Řezy mozkem – nepostradatelný průvodce anatomickými detaily a diagnostickými metodami – spojuje makroskopické, mikroskopické a MRI řezy mozku do jednoho uceleného a systematického celku. Poskytuje jasné vizuální pochopení toho, jak mozek vypadá a jak jej lze interpretovat prostřednictvím moderních diagnostických metod, jejichž neustálý vývoj vyžaduje od studentů a lékařů, aby se orientovali nejen v zevní anatomii mozku, ale i v řezech. Vyšetření magnetickou rezonancí a výpočetní tomografií patří dnes mezi běžné metody a jednoznačným předpokladem k jejich interpretaci je znalost anatomie. Nejsrozumitelnější přístup k anatomii mozku – bohatá obrazová dokumentace umožňuje vidět mozek při pitvě nebo ve špičkově vybavené laboratoři. Ať už jde o frontální, transverzální nebo sagitální roviny, vše je zachyceno a doplněno přesnými schématy, která pomáhají pochopit polohu a orientaci jednotlivých řezů. Zdařilá kombinace teorie a praxe – text poskytuje vše, co je třeba pro interpretaci snímků z MRI nebo CT vyšetření, od makroskopické stavby až po mikroskopické detaily. Exkluzivní mikroskopické řezy – unikátní soubor mikroskopických řezů umožňuje porozumět hlavním strukturám, což pomáhá identifikaci jak pod mikroskopem, tak na diagnostických snímcích. Přehledná orientace díky současné anatomické nomenklatuře. Předpokladem k účelnému využití této publikace je základní znalost anatomie CNS, neboť atlas se soustředí výhradně na obrazovou dokumentaci. Kniha je určena studentům lékařství, neurologům, neuroradiologům, ale i dalším odborníkům, které fascinuje mozek a chtějí si rozšířit znalosti o jeho stavbě.

• MeSH
• magnetická rezonanční tomografie MeSH
• mikroskopie MeSH
• mozek anatomie a histologie   diagnostické zobrazování   MeSH
• neuroanatomie MeSH
• Publikační typ
• atlasy MeSH

• Konspekt
• Anatomie člověka a srovnávací anatomie
• NLK Obory
• neurovědy
• radiologie, nukleární medicína a zobrazovací metody

Rentgenová výpočetní mikrotomografie (mikroCT) představuje moderní zobrazovací technologii s vysokým rozlišením umožňující detailní analýzu zobrazovaného vzorku. Nabízí jedinečný pohled na trojrozměrnou architekturu díky rozlišení na pomezí makroskopického a histologického zobrazení. V oblasti anatomické patologie mikroCT nachází uplatnění zejména při morfometrické analýze nádorů, hodnocení resekčních okrajů chirurgických vzorků či detekci metastáz v lymfatických uzlinách. Kombinace mikroCT s tradičními histopatologickými technikami a s využitím digitální 3D rekonstrukce otevírá nové možnosti při analýze komplexních patologických procesů. Přestože je tato metoda zatím převážně využívána ve výzkumu, její klinický potenciál je značný. Mezi hlavní přednosti patří neinvazivní zobrazení a možnost integrace s digitální patologií a nástroji umělé inteligence. Hlavními limitacemi v současné době zůstávají potřeba kontrastování vzorků, monochromatická povaha obrazu a vysoká radiační zátěž. Pokrok v technologickém vývoji však může tyto překážky překonat a umožnit širší využití mikroCT v rutinní klinické diagnostice. Tento článek představuje technologii mikroCT a její diagnostický potenciál v patologii, přibližuje její aplikace, výhody a omezení, a nabízí vhled do budoucí perspektivy jejího využití.

X-ray microtomography (microCT) represents a modern high-resolution imaging technology enabling detailed analysis of the tissue. It offers a unique perspective on three-dimensional architecture, bridging the gap between macroscopic and histological imaging. In anatomical pathology, microCT is particularly utilized for morphometric tumor analysis, evaluation of surgical specimen resection margins, and detection of metastases in lymph nodes. The combination of microCT with traditional histopathological techniques, and with digital 3D reconstructions, opens new avenues for analyzing complex pathological processes. Although this method is currently used in research, its clinical potential is significant. Key advantages include non-invasive imaging and the ability to be integrated with digital pathology and artificial intelligence tools. Current limitations include the need for sample contrast enhancement, the monochromatic nature of the images, and high radiation exposure. Advances in technological development, however, may overcome these barriers and enable the broader adoption of microCT in routine clinical diagnostics. This article explores the diagnostic potential of microCT in pathology, highlighting its applications, advantages, and limitations, while offering insights into current capabilities and future perspectives of this technology.

• MeSH
• biopsie trendy   MeSH
• lidé MeSH
• mikroskopie přístrojové vybavení   trendy   MeSH
• patologie * přístrojové vybavení   trendy   MeSH
• rentgenová mikrotomografie * metody   trendy   MeSH
• zobrazování trojrozměrné trendy   MeSH
• Check Tag
• lidé MeSH

Recent advances in optical sensing technologies underpin the development of high-performance, surface-sensitive analytical tools capable of reliable and precise detection of molecular targets in complex biological media in non-laboratory settings. Optical fibre sensors guide light to and from a region of interest, enabling sensitive measurements of localized environments. This positions optical fibre sensors as a highly promising technology for a wide range of biochemical and healthcare applications. However, their performance in real-world biological media is often limited by the absence of robust post-modification strategies that provide both high biorecognition and antifouling capabilities. In this study, we present the proof-of-concept antifouling and biorecognition performance of a polymer brush nano-coating synthesized at the sensing region of optical fibre long-period grating (LPG) sensors. Using a newly developed antifouling terpolymer brush (ATB) composed of carboxybetaine methacrylamide, sulfobetaine methacrylamide, and N-(2-hydroxypropyl)methacrylamide, we achieve state-of-the-art antifouling properties. The successful on-fibre ATB synthesis is confirmed through scanning electron microscopy (SEM), fluorescence microscopy, and label-free bio-detection experiments based on antibody-functionalized ATB-coated LPG optical fibres. Despite the challenges in handling optical fibres during polymerization, the resulting nano-coating retains its remarkable antifouling properties upon exposure to blood plasma and enables biorecognition element functionalization. These capabilities are demonstrated through the detection of IgG in buffer and diluted blood plasma using anti-IgG-functionalized ATB-coated sensing regions of LPG fibres in both label-based (fluorescence) and label-free real-time detection experiments. The results show the potential of ATB-coated LPG fibres for use in analytical biosensing applications.

• MeSH
• akrylamidy chemie   MeSH
• biosenzitivní techniky * přístrojové vybavení   MeSH
• bioznečištění * prevence a kontrola   MeSH
• nanostruktury chemie   MeSH
• optická vlákna * MeSH
• polymery chemie   MeSH
• Publikační typ
• časopisecké články MeSH

Quantitative genomic mapping of DNA damage may provide insights into the underlying mechanisms of damage and repair. Sequencing based approaches are bound to the limitations of PCR amplification bias and read length which hamper both the accurate quantitation of damage events and the ability to map them to structurally complex genomic regions. Optical Genome mapping in arrays of parallel nanochannels allows physical extension and genetic profiling of millions of long genomic DNA fragments, and has matured to clinical utility for characterization of complex structural aberrations in cancer genomes. Here we present a new mapping modality, Repair-Assisted Damage Detection - Optical Genome Mapping (RADD-OGM), a method for single-molecule level mapping of DNA damage on a genome-wide scale. Leveraging ultra-long reads to assemble the complex structure of a sarcoma cell-line genome, we mapped the genomic distribution of oxidative DNA damage, identifying regions more susceptible to DNA oxidation. We also investigated DNA repair by allowing cells to repair chemically induced DNA damage, pinpointing locations of concentrated repair activity, and highlighting variations in repair efficiency. Our results showcase the potential of the method for toxicogenomic studies, mapping the effect of DNA damaging agents such as drugs and radiation, as well as following specific DNA repair pathways by selective induction of DNA damage. The facile integration with optical genome mapping enables performing such analyses even in highly rearranged genomes such as those common in many cancers, a challenging task for sequencing-based approaches.

• MeSH
• bromičnany toxicita   MeSH
• lidé MeSH
• mapování chromozomů * přístrojové vybavení   metody   MeSH
• mikrofluidní analytické techniky * přístrojové vybavení   metody   MeSH
• nádorové buněčné linie MeSH
• nanotechnologie * přístrojové vybavení   metody   MeSH
• oprava DNA genetika   MeSH
• oxidační stres účinky léků   genetika   MeSH
• poškození DNA * genetika   MeSH
• regulace genové exprese MeSH
• stanovení celkové genové exprese MeSH
• toxikogenetika * přístrojové vybavení   metody   MeSH
• variabilita počtu kopií segmentů DNA MeSH
• zobrazení jednotlivé molekuly * přístrojové vybavení   metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

This study aimed to compare the fascicular anatomy of upper limb nerves visualized using in situ high-resolution ultrasound (HRUS) with ex vivo imaging modalities, namely, magnetic resonance microscopy (MRM), histological cross-sections (HCS), and optical projection tomography (OPT). The median, ulnar, and superficial branch of radial nerve (n = 41) were visualized in 14 cadaveric upper limbs using 22-MHz HRUS. Subsequently, the nerves were excised, imaged with different microscopic techniques, and their morphometric properties were compared. HRUS accurately differentiated 51-74% of fascicles, while MRM detected 87-92% of fascicles when compared to the referential HCS. Among the compared modalities, HRUS demonstrated the smallest fascicular ratios and fascicular cross-sectional areas, but the largest nerve cross-sectional areas. The probability of a fascicle depicted on HRUS representing a cluster of multiple fascicles on the referential HCS increased with the fascicular size, with some differences observed between the larger median and ulnar nerves and the smaller radial nerves. Accordingly, HRUS fascicle differentiation necessitates cautious interpretation, as larger fascicles are more likely to represent clusters. Although HCS is considered the reference modality, alterations in nerve cross-sectional areas or roundness during sample processing should be acknowledged.

• MeSH
• horní končetina * inervace   diagnostické zobrazování   MeSH
• lidé středního věku MeSH
• lidé MeSH
• magnetická rezonanční tomografie metody   MeSH
• mikroskopie metody   MeSH
• mrtvola MeSH
• nervus medianus diagnostické zobrazování   MeSH
• nervus radialis * diagnostické zobrazování   anatomie a histologie   MeSH
• nervus ulnaris * diagnostické zobrazování   anatomie a histologie   MeSH
• senioři MeSH
• ultrasonografie * metody   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The GPCR signalling cascade is a key pathway responsible for the signal transduction of a multitude of physical and chemical stimuli, including light, odorants, neurotransmitters and hormones. Understanding the structural and functional properties of the GPCR cascade requires direct observation of signalling processes in high spatial and temporal resolution, with minimal perturbation to endogenous systems. Optical microscopy and spectroscopy techniques are uniquely suited to this purpose because they excel at multiple spatial and temporal scales and can be used in living objects. Here, we review recent developments in microscopy and spectroscopy technologies which enable new insights into GPCR signalling. We focus on advanced techniques with high spatial and temporal resolution, single-molecule methods, labelling strategies and approaches suitable for endogenous systems and large living objects. This review aims to assist researchers in choosing appropriate microscopy and spectroscopy approaches for a variety of applications in the study of cellular signalling. LINKED ARTICLES: This article is part of a themed issue Complexity of GPCR Modulation and Signaling (ERNST). To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v182.14/issuetoc.

• MeSH
• lidé MeSH
• mikroskopie * metody   MeSH
• receptory spřažené s G-proteiny * chemie   metabolismus   MeSH
• signální transdukce MeSH
• spektrální analýza * metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

STUDY QUESTION: Can oocyte functionality be assessed by observing changes in their intracytoplasmic lipid droplets (LDs) profiles? SUMMARY ANSWER: Lipid profile changes can reliably be detected in human oocytes; lipid changes are linked with maternal age and impaired developmental competence in a mouse model. WHAT IS KNOWN ALREADY: In all cellular components, lipid damage is the earliest manifestation of oxidative stress (OS), which leads to a cascade of negative consequences for organelles and DNA. Lipid damage is marked by the accumulation of LDs. We hypothesized that impaired oocyte functionality resulting from aging and associated OS could be assessed by changes in LDs profile, hereafter called lipid fingerprint (LF). STUDY DESIGN, SIZE, DURATION: To investigate if it is possible to detect differences in oocyte LF, we subjected human GV-stage oocytes to spectroscopic examinations. For this, a total of 48 oocytes derived from 26 young healthy women (under 33 years of age) with no history of infertility, enrolled in an oocyte donation program, were analyzed. Furthermore, 30 GV human oocytes from 12 women were analyzed by transmission electron microscopy (TEM). To evaluate the effect of oocytes' lipid profile changes on embryo development, a total of 52 C57BL/6 wild-type mice and 125 Gnpat+/- mice were also used. PARTICIPANTS/MATERIALS, SETTING, METHODS: Human oocytes were assessed by label-free cell imaging via coherent anti-Stokes Raman spectroscopy (CARS). Further confirmation of LF changes was conducted using spontaneous Raman followed by Fourier transform infrared (FTIR) spectroscopies and TEM. Additionally, to evaluate whether LF changes are associated with developmental competence, mouse oocytes and blastocysts were evaluated using TEM and the lipid dyes BODIPY and Nile Red. Mouse embryonic exosomes were evaluated using flow cytometry, FTIR and FT-Raman spectroscopies. MAIN RESULTS AND THE ROLE OF CHANCE: Here we demonstrated progressive changes in the LF of oocytes associated with the woman's age consisting of increased LDs size, area, and number. LF variations in oocytes were detectable also within individual donors. This finding makes LF assessment a promising tool to grade oocytes of the same patient, based on their quality. We next demonstrated age-associated changes in oocytes reflected by lipid peroxidation and composition changes; the accumulation of carotenoids; and alterations of structural properties of lipid bilayers. Finally, using a mouse model, we showed that LF changes in oocytes are negatively associated with the secretion of embryonic exosomes prior to implantation. Deficient exosome secretion disrupts communication between the embryo and the uterus and thus may explain recurrent implantation failures in advanced-age patients. LIMITATIONS, REASONS FOR CAUTION: Due to differences in lipid content between different species' oocytes, the developmental impact of lipid oxidation and consequent LF changes may differ across mammalian oocytes. WIDER IMPLICATIONS OF THE FINDINGS: Our findings open the possibility to develop an innovative tool for oocyte assessment and highlight likely functional connections between oocyte LDs and embryonic exosome secretion. By recognizing the role of oocyte LF in shaping the embryo's ability to implant, our original work points to future directions of research relevant to developmental biology and reproductive medicine. STUDY FUNDING/COMPETING INTEREST(S): This research was funded by National Science Centre of Poland, Grants: 2021/41/B/NZ3/03507 and 2019/35/B/NZ4/03547 (to G.E.P.); 2022/44/C/NZ4/00076 (to M.F.H.) and 2019/35/N/NZ3/03213 (to Ł.G.). M.F.H. is a National Agency for Academic Exchange (NAWA) fellow (GA ULM/2019/1/00097/U/00001). K.F. is a Diamond Grant fellow (Ministry of Education and Science GA 0175/DIA/2019/28). The open-access publication of this article was funded by the Priority Research Area BioS under the program "Excellence Initiative - Research University" at the Jagiellonian University in Krakow. The authors declare no competing interest. TRIAL REGISTRATION NUMBER: N/A.

• MeSH
• dospělí MeSH
• embryonální vývoj fyziologie   MeSH
• lidé MeSH
• lipidová tělíska metabolismus   MeSH
• metabolismus lipidů MeSH
• myši inbrední C57BL * MeSH
• myši MeSH
• oocyty * metabolismus   MeSH
• oxidační stres MeSH
• Ramanova spektroskopie MeSH
• stárnutí metabolismus   MeSH
• transmisní elektronová mikroskopie MeSH
• věk matky MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH