PURPOSE: Indocyanine green (ICG) fluorescence imaging enhances biliary visualization during pediatric laparoscopic cholecystectomy (LC), helping to identify anatomical variants and prevent bile duct injury. Standard pediatric recommendations suggest ICG administration 16-24 h preoperatively; however, this may be impractical. This study aims to evaluate the safety and effectiveness of short-interval ICG administration. METHODS: A prospective single-center study (October 2024-June 2025) included pediatric LC patients receiving intravenous Verdye® preoperatively. Visualization of extrahepatic biliary anatomy was assessed intraoperatively using a 5-point Likert scale, HELPFUL (usefulness), and DISTURBED (liver background interference) scores. Data included indication, ICG timing, operative time, and complications according to the Clavien-Dindo classification (C-D). RESULTS: Eleven patients (64% female), median age 14 years (IQR 12,7-15,7) and median weight 65,5 kg (IQR 46,5-80), were included. Five had BMI > 25 kg/m2; five (46%) underwent preoperative ERCP. ICG (median dose 0.34 mg/kg) was administered a median of 225 min before surgery. Median operative time was 65 min (IQR 58-68). Median Likert score was 5; HELPFUL 3; DISTURBED 1. No ICG-related or C-D complications occurred. CONCLUSION: Short-interval ICG administration was safe, feasible, and effective in enhancing biliary visualization during pediatric LC. This approach was well-tolerated and provided high-quality imaging without complications.

• Klíčová slova
• Biliary visualization, Fluorescence imaging, Indocyanine green, Pediatric cholecystectomy, Short-interval ICG administration,
• MeSH
• barvicí látky aplikace a dávkování   škodlivé účinky   MeSH
• časové faktory MeSH
• chirurgie s pomocí počítače * metody   MeSH
• cholecystektomie laparoskopická * škodlivé účinky   metody   MeSH
• délka operace MeSH
• délka pobytu MeSH
• dítě MeSH
• indokyanová zeleň * aplikace a dávkování   škodlivé účinky   MeSH
• injekce intravenózní MeSH
• lidé MeSH
• mladiství MeSH
• optické zobrazování * škodlivé účinky   metody   MeSH
• pooperační komplikace epidemiologie   MeSH
• předoperační péče škodlivé účinky   metody   MeSH
• prospektivní studie MeSH
• studie proveditelnosti MeSH
• žlučové ústrojí diagnostické zobrazování   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• Názvy látek
• barvicí látky MeSH
• indokyanová zeleň * MeSH

Fluorescence Lifetime Imaging Microscopy (FLIM) of endogenous fluorophores has recently emerged as a powerful, marker-free, and non-invasive tool for investigating cellular metabolism. This cutting-edge imaging technique provides valuable insights into cellular energy states by measuring the fluorescence lifetimes of intrinsically fluorescent redox cofactors. The lifetimes of these cofactors reflect their binding states to enzymes, thus indicating enzymatic activity within specific metabolic pathways. As a result, FLIM can help to reveal the overall redox status of the cell and, to some extent, shifts between oxidative phosphorylation and glycolysis. The application of FLIM in metabolic research has shown significant progress across a diverse range of pathological contexts, including cancer, diabetes, neurodegenerative disorders, and various forms of cardiopathology.The aim of this mini-review is to introduce the methodology of NAD(P)H and FAD/FMN FLIM, outline its underlying principles, and demonstrate its ability to reveal changes in cellular metabolism. Additionally, this mini-review highlights FLIM's potential for understanding cellular redox states, detecting metabolic shifts in various disease models, and contributing to the development of therapeutic strategies.

• Klíčová slova
• FAD, FLIM, FLIRR, NAD(P)H, Redox state,
• MeSH
• flavinadenindinukleotid metabolismus   MeSH
• fluorescenční barviva * MeSH
• fluorescenční mikroskopie metody   MeSH
• lidé MeSH
• NADP metabolismus   MeSH
• neurodegenerativní nemoci metabolismus   MeSH
• optické zobrazování metody   MeSH
• oxidace-redukce MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• flavinadenindinukleotid MeSH
• fluorescenční barviva * MeSH
• NADP MeSH

INTRODUCTION: Macular pigment plays an important role in the reduction of oxidative stress and in preventing retinal diseases. Quick and easy measurements of the macular pigment are essential in both clinical and research settings. Dual wavelength fundus auto-fluorescence seems to be the optimal method. This study aims to investigate changes in fundus autofluorescence in patients taking daily lutein oral supplements and develop image processing methods for follow-up evaluations of the images. METHODS: New devices allow us to examine fundus autofluorescence using both blue and green excitation wavelengths. This allows detection of the amount of macular pigment by subtracting these two images because the yellow pigment particles absorb blue wavelengths. We determined daily dose of 25 mg of lutein and 3 mg of zeaxanthin. Patients were followed up for 15 months at 3-month intervals. RESULTS: During our 15-month study, we observed a positive trend in pixel lightness values, suggesting an increase in macular pigments in the foveal area. In all patients taking daily lutein supplements, the foveal index significantly increased after six months, with a median change of 0.081. We did not observe a significant change after the first three months (0.006) and only a small change between the 6th and 12th-month visits (0.012). CONCLUSION: With appropriate patients and procedures for capturing autofluorescence images, this is a valuable technique for macular pigment evaluation in follow-up examinations using software image post-processing and analysis with commonly available hardware. To put this into everyday practice, developing tools to automate the assessment is necessary.

• Klíčová slova
• auto-fluorescence, lutein, macula, macular pigment, retina,
• MeSH
• dospělí MeSH
• fundus oculi MeSH
• lidé středního věku MeSH
• lidé MeSH
• lutein * aplikace a dávkování   MeSH
• makulární pigment * analýza   metabolismus   MeSH
• optické zobrazování * metody   MeSH
• potravní doplňky MeSH
• senioři MeSH
• zeaxanthiny aplikace a dávkování   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• lutein * MeSH
• makulární pigment * MeSH
• zeaxanthiny MeSH

AIMS: To present a new method of dynamic Purkinje-metry and to verify it by comparison with a commercially available anterior segment optical coherence tomography CASIA2. PATIENTS AND METHODS: A dynamic Purkinje-meter with a movable fixation target was assembled. A coaxial circular pattern formed by infrared LEDs was projected onto the eye and evoked Purkinje images (1st, 3rd, 4th = P1, P3, P4). The measurement was performed on 29 eyes with an implanted toric IOL (intraocular lens), under mydriatic conditions, with reference to the visual axis. The IOL tilt was calculated from the position of a fixation target at the moment of P3 and P4 superposition. The IOL decentration was determined based on the relative position of P1 during on-axis fixation and of P3 and P4 superposition during off-axis fixation. A custom-developed software was used for distance measurements. Using CASIA2, the IOL position was fully calculated by the device. RESULTS: The mean absolute difference between CASIA2 and Purkinje-meter values was 0.6° ± 0.4° for the tilt magnitude and 10° ± 10° for the tilt direction, and 0.11 mm ± 0.08 mm for the decentration magnitude and 16° ± 14° for the decentration direction. There was no statistically significant difference between the values determined by the two methods for the tilt and decentration direction. The differences were statistically significant for the tilt and decentration magnitude. CONCLUSION: The values of IOL tilt and decentration direction are similar for both devices. The values of IOL tilt and decentration magnitude measured by Purkinje-meter are higher than those from CASIA2, but overall, they correspond to the values presented in other published studies.

• Klíčová slova
• CASIA2, Purkinje images, anterior segment optical coherence tomography, intraocular lens decentration, intraocular lens position, intraocular lens tilt,
• MeSH
• algoritmy MeSH
• lidé středního věku MeSH
• lidé MeSH
• nitrooční čočky * MeSH
• optická koherentní tomografie MeSH
• optické zobrazování * přístrojové vybavení   metody   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky MeSH
• srovnávací studie MeSH

Fluorescence-based contrast agents enable real-time detection of solid tumors and their neovasculature, making them ideal for use in image-guided surgery. Several agents have entered late-stage clinical trials or secured FDA approval, suggesting they are likely to become the standard of care in cancer surgeries. One of the key parameters to optimize in contrast agents is molecular size, which dictates much of the pharmacokinetic and pharmacodynamic properties of the agent. Here, we describe the development of a class of protease-activated quenched fluorescent probes in which a N-(2-hydroxypropyl)methacrylamide copolymer is used as the primary scaffold. This copolymer core provides a high degree of probe modularity to generate structures that cannot be achieved with small molecules and peptide probes. We used a previously validated cathepsin substrate and evaluated the effects of length and type of linker, as well as the positioning of the fluorophore/quencher pair on the polymer core. We found that the polymeric probes could be optimized to achieve increased overall signal and tumor-to-background ratios compared to the reference small molecule probe. Our results also revealed multiple structure-activity relationship trends that can be used to design and optimize future optical imaging probes. Furthermore, they confirm that a hydrophilic polymer is an ideal scaffold for use in optical imaging contrast probes, allowing a highly modular design that enables efficient optimization to maximize probe accumulation and overall biodistribution properties.

• Klíčová slova
• HPMA copolymer, cancer, fluorescence, iBody, imaging, protease,
• MeSH
• akrylamidy chemie   MeSH
• fluorescenční barviva * chemie   chemická syntéza   MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory * diagnostické zobrazování   MeSH
• optické zobrazování metody   MeSH
• polymery chemie   MeSH
• proteasy metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• akrylamidy MeSH
• fluorescenční barviva * MeSH
• N-(2-hydroxypropyl)methacrylamide MeSH Prohlížeč
• polymery MeSH
• proteasy MeSH

Herein, we report the development of a macromolecular multifunctional imaging tool for biological investigations, which is comprised of an N-(2-hydroxypropyl)methacrylamide backbone, iridium-based luminescent probe, glutamate carboxypeptidase II (GCPII) targeting ligand, and biotin affinity tag. The iridium luminophore is a tris-cyclometalated complex based on [Ir(ppy)3] with one of its 2-phenylpyridine ligands functionalized to allow conjugation. Synthesized macromolecular probes differed in the structure of the polymer and content of the iridium complex. The applicability of the developed imaging tools has been tested in flow cytometry (FACS) based assay, laser confocal microscopy, and fluorescence lifetime imaging microscopy (FLIM). The FACS analysis has shown that the targeted iBodies containing the iridium luminophore exhibit selective labelling of GCPII expressing cells. This observation was also confirmed in the imaging experiments with laser confocal microscopy. The FLIM experiment has shown that the iBodies with the iridium label exhibit a lifetime greater than 100 ns, which distinguishes them from typically used systems labelled with organic fluorophores exhibiting short fluorescence lifetimes. The results of this investigation indicate that the system exhibits interesting properties, which supports the development of additional biological tools utilizing the key components (iridium complexes, iBody concept), primarily focusing on the longer lifetime of the iridium emitter.

• Klíčová slova
• HPMA, iBody, imaging, iridium complex,
• MeSH
• fluorescenční barviva chemie   MeSH
• fluorescenční mikroskopie metody   MeSH
• iridium * chemie   MeSH
• konfokální mikroskopie * MeSH
• lidé MeSH
• optické zobrazování metody   MeSH
• polymery * chemie   MeSH
• průtoková cytometrie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

A novel method for semiautomated assessment of directions of collagen fibers in soft tissues using histological image analysis is presented. It is based on multiple rotated images obtained via polarized light microscopy without any additional components, i.e., with just two polarizers being either perpendicular or nonperpendicular (rotated). This arrangement breaks the limitation of 90° periodicity of polarized light intensity and evaluates the in-plane fiber orientation over the whole 180° range accurately and quickly. After having verified the method, we used histological specimens of porcine Achilles tendon and aorta to validate the proposed algorithm and to lower the number of rotated images needed for evaluation. Our algorithm is capable to analyze 5·105 pixels in one micrograph in a few seconds and is thus a powerful and cheap tool promising a broad application in detection of collagen fiber distribution in soft tissues.

• MeSH
• Achillova šlacha metabolismus   MeSH
• algoritmy MeSH
• extracelulární matrix metabolismus   MeSH
• kolagen metabolismus   MeSH
• mikroskopie metody   MeSH
• optické zobrazování metody   MeSH
• počítačové zpracování obrazu metody   MeSH
• polarizační mikroskopie metody   MeSH
• prasata MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kolagen MeSH

Upconverting luminescent lanthanide-doped nanoparticles (UCNP) belong to promising new materials that absorb infrared light able to penetrate in the deep tissue level, while emitting photons in the visible or ultraviolet region, which makes them favorable for bioimaging and cell labeling. Here, we have prepared upconverting NaYF4:Yb,Er@NaYF4:Nd core-shell nanoparticles, which were coated with copolymers of N,N-dimethylacrylamide (DMA) and 2-(acryloylamino)-2-methylpropane-1-sulfonic acid (AMPS) or tert-butyl [2-(acryloylamino)ethyl]carbamate (AEC-Boc) with negative or positive charges, respectively. The copolymers were synthesized by a reversible addition-fragmentation chain transfer (RAFT) polymerization, reaching Mn ~ 11 kDa and containing ~ 5 mol% of reactive groups. All copolymers contained bisphosphonate end-groups to be firmly anchored on the surface of NaYF4:Yb,Er@NaYF4:Nd core-shell nanoparticles. To compare properties of polymer coatings, poly(ethylene glycol)-coated and neat UCNP were used as a control. UCNP with various charges were then studied as labels of carcinoma cells, including human hepatocellular carcinoma HepG2, human cervical cancer HeLa, and rat insulinoma INS-1E cells. All the particles proved to be biocompatible (nontoxic); depending on their ξ-potential, the ability to penetrate the cells differed. This ability together with the upconversion luminescence are basic prerequisites for application of particles in photodynamic therapy (PDT) of various tumors, where emission of nanoparticles in visible light range at ~ 650 nm excites photosensitizer.

• MeSH
• akrylamidy chemie   MeSH
• buňky Hep G2 MeSH
• fluorescenční barviva chemie   MeSH
• fluoridy chemie   MeSH
• HeLa buňky MeSH
• lidé MeSH
• nádory diagnostické zobrazování   MeSH
• nanočástice chemie   MeSH
• optické zobrazování metody   MeSH
• ytrium chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• akrylamidy MeSH
• fluorescenční barviva MeSH
• fluoridy MeSH
• poly(N,N-dimethylacrylamide) MeSH Prohlížeč
• sodium yttriumtetrafluoride MeSH Prohlížeč
• ytrium MeSH

The authors present their contribution to the improvement of methods suitable for the detection of the freezing and thawing damage of cells of cryopreserved venous grafts used for lower limb revascularization procedures. They studied the post-thaw viability of cells of the wall of cryopreserved venous grafts (CVG) immediately after thawing and after 24 and 48 h culture at +37 °C in two groups of six CVG selected randomly for slow thawing in the refrigerator and rapid thawing in a water bath at +37 °C. The grafts were collected from multi-organ and tissue brain-dead donors, cryopreserved, and stored in a liquid nitrogen vapor phase for five years. The viability was assessed from tissue slices obtained by perpendicular and longitudinal cuts of the thawed graft samples using in situ staining with fluorescence vital dyes. The mean and median immediate post-thaw viability values above 70% were found in using both thawing protocols and both types of cutting. The statistically significant decline in viability after the 48-h culture was observed only when using the slow thawing protocol and perpendicular cutting. The possible explanation might be the "solution effect damage" during slow thawing, which caused a gentle reduction in the graft cellularity. The possible influence of this phenomenon on the immunogenicity of CVG should be the subject of further investigations.

• Klíčová slova
• Thawing method, cell viability, confocal microscopy, cryopreservation, fluorescence vital dyes, vascular allograft,
• MeSH
• alografty diagnostické zobrazování   účinky léků   MeSH
• apoptóza účinky léků   MeSH
• dárci tkání MeSH
• dimethylsulfoxid farmakologie   MeSH
• fluorescenční barviva * MeSH
• konfokální mikroskopie metody   MeSH
• kryoprezervace metody   MeSH
• kryoprotektivní látky farmakologie   MeSH
• lidé MeSH
• optické zobrazování metody   MeSH
• transplantace cév metody   MeSH
• vena femoralis diagnostické zobrazování   účinky léků   MeSH
• vena saphena diagnostické zobrazování   účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• zmrazování * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• dimethylsulfoxid MeSH
• fluorescenční barviva * MeSH
• kryoprotektivní látky MeSH

Surgery is an efficient way to treat localized prostate cancer (PCa), however, it is challenging to demarcate rapidly and accurately the tumor boundary intraoperatively, as existing tumor detection methods are seldom performed in real-time. To overcome those limitations, we develop a fluorescent molecular rotor that specifically targets the prostate-specific membrane antigen (PSMA), an established marker for PCa. The probes have picomolar affinity (IC50 = 63-118 pM) for PSMA and generate virtually instantaneous onset of robust fluorescent signal proportional to the concentration of the PSMA-probe complex. In vitro and ex vivo experiments using PCa cell lines and clinical samples, respectively, indicate the utility of the probe for biomedical applications, including real-time monitoring of endocytosis and tumor staging. Experiments performed in a PCa xenograft model reveal suitability of the probe for imaging applications in vivo.

• MeSH
• antigeny povrchové chemie   metabolismus   MeSH
• buňky PC-3 MeSH
• endocytóza MeSH
• fluorescenční spektrometrie metody   MeSH
• glutamátkarboxypeptidasa II chemie   metabolismus   MeSH
• lidé MeSH
• molekulární modely MeSH
• molekulární sondy chemie   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši nahé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory prostaty diagnóza   metabolismus   MeSH
• optické zobrazování metody   MeSH
• proteinové domény MeSH
• transplantace heterologní MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• antigeny povrchové MeSH
• FOLH1 protein, human MeSH Prohlížeč
• glutamátkarboxypeptidasa II MeSH
• molekulární sondy MeSH