Innovative Imaging Techniques Research Articles

Cardiovascular comorbidities in psoriatic arthritis: state of the art.

Psoriatic arthritis (PsA) is a complex multi-system immune-mediated condition, characterised by a high comorbidity burden, one of the most prevalent of which is cardiovascular disease (CVD), affecting up to 80% of patients. This narrative review explores the current understanding of cardiovascular comorbidities in PsA, focusing on mechanistic pathways, risk assessment, and the impact of treatment choices on cardiovascular health. Here, we outline the role of inflammatory cytokines, immune system dysregulation, and genetic predispositions in PsA, not only as drivers of musculoskeletal manifestations but also atherosclerosis and endothelial dysfunction, giving rise to cardiovascular pathology. Given these insights, accurately assessing and predicting cardiovascular risk in PsA patients is a critical challenge. This review evaluates traditional risk calculators as well as innovative biomarkers and imaging techniques, emphasising their utility and limitations in capturing the true cardiovascular risk profile of PsA patients. There are multiple complexities surrounding the treatment of PsA in the context of concurrent CVD, and therapeutic choices must carefully balance efficacy in managing PsA symptoms with the potential cardiovascular implications. A multidisciplinary approach, integrating dermatological, rheumatological, and cardiological perspectives, amongst others, to optimise patient outcomes, is key. Overall, a heightened clinical awareness and research focus on cardiovascular comorbidities in PsA is warranted, aiming to refine risk assessment strategies and therapeutic interventions that holistically address the multifaceted needs of patients with PsA.

Label-Free Non-Contact Vascular Imaging using Photon Absorption Remote Sensing.

Functional vascular imaging is a critical method for early detection and prevention of disease. Established non-contact vascular imaging techniques capture predominantly structural information. In this study, a novel non-contact label-free in vivo Photon Absorption Remote Sensing (PARS) microscope is developed for structural and functional vascular imaging. The presented in vivo PARS microscope captures the endogenous absorption of green (532nm) light to form a complete picture of vasculature and surrounding tissues. Imaging system repeatability is enhanced through robust transient absorption signal extraction, and state-of-the-art real-time alignment methods. Detailed imaging of vascular structure is demonstrated through in vivo microscopy of two established animal models: mouse ear and chicken embryo. Preliminary functional contrast is realized through video rate imaging of red blood cell dynamics in the capillary networks of chicken embryos. The presented in vivo PARS microscope successfully captures detailed structural and functional vascular contrast. This innovative non-contact label-free imaging technique holds promise as a tool for preventative medical care, as functional change often precedes structural change.

Biochemical Detection of Capsid in the Nucleus During HIV-1 Infection.

Recent evidence has shown that uncoating and reverse transcription precede nuclear import. These recent breakthroughs have been made possible through the development of innovative biochemical and imaging techniques. This method outlines the biochemical assay used for detecting the presence of the HIV-1 core in the nuclear compartment. In this procedure, human cells are infected with HIV-1NL4-3, with or without the inclusion of PF74, a small molecule that inhibits core entry into the nuclear compartment. Subsequently, cells are separated into cytosolic and nuclear fractions. To assess whether the capsid protein has reached the nuclear compartment, cytosolic and nuclear fractions are subjected to Western blot analysis, utilizing antibodies specific to the HIV-1 capsid protein p24. To validate the true origin of these fractions, Western blot analysis employing antibodies against cytosolic and nuclear markers arealso performed. In summary, this assay provides a reliable and efficient means to detect the presence of the HIV-1 capsid protein in the nucleus during infection under various conditions.

X-ray-induced acoustic computed tomography and its applications in biomedicine.

X-ray-induced acoustic computed tomography (XACT) offers a promising approach to biomedical imaging, leveraging X-ray absorption contrast. It overcomes the shortages of traditional X-ray, allowing for more advanced medical imaging. The review focuses on the significance and draws onto the potential applications of XACT to demonstrate it as an innovative imaging technique. This review navigates the expanding landscape of XACT imaging within the biomedical sphere. Integral topics addressed encompass the refinement of imaging systems and the advancement in image reconstruction algorithms. The review particularly emphasizes XACT's significant biomedical applications. Key uses, such as breast imaging, bone density maps for osteoporosis, and X-ray molecular imaging, are highlighted to demonstrate the capability of XACT. A unique niche for XACT imaging is its application in in vivo dosimetry during radiotherapy, which has been validated on patients. Because of its unique property, XACT has great potential in biomedicine and non-destructive testing. We conclude by casting light on potential future avenues in this promising domain.

Innovative Imaging and Analysis Techniques for Quantifying Spalling Repair Materials in Concrete Pavements

Traditional spalling repair on concrete pavement roads is labor-intensive. It involves traffic blockages and the manual calculation of repair areas, leading to time-consuming processes with potential discrepancies. This study used a line scan camera to photograph road surface conditions to analyze spalling without causing traffic blockage in an indoor setting. By using deep learning algorithms, specifically a region-based convolutional neural network (R-CNN) in the form of the Mask R-CNN algorithm, the system detects spalling and calculates its area. The program processes data based on the Federal Highway Administration (FHWA) spalling repair standards. Accuracy was assessed using root mean square error (RMSE) and Pearson correlation coefficient (PCC) via comparisons with actual field calculations. The RMSE values were 0.0137 and 0.0167 for the minimum and maximum repair areas, respectively, showing high accuracy. The PCC values were 0.987 and 0.992, indicating a strong correlation between the actual and calculated repair areas, confirming the high calculation accuracy of the method.

An mTurq2-Col4a1 mouse model allows for live visualization of mammalian basement membrane development.

Basement membranes (BMs) are specialized sheets of extracellular matrix that underlie epithelial and endothelial tissues. BMs regulate the traffic of cells and molecules between compartments, and participate in signaling, cell migration, and organogenesis. The dynamics of mammalian BMs, however, are poorly understood, largely due to a lack of models in which core BM components are endogenously labeled. Here, we describe the mTurquoise2-Col4a1 mouse in which we fluorescently tag collagen IV, the main component of BMs. Using an innovative planar-sagittal live imaging technique to visualize the BM of developing skin, we directly observe BM deformation during hair follicle budding and basal progenitor cell divisions. The BM's inherent pliability enables dividing cells to remain attached to and deform the BM, rather than lose adhesion as generally thought. Using FRAP, we show BM collagen IV is extremely stable, even during periods of rapid epidermal growth. These findings demonstrate the utility of the mTurq2-Col4a1 mouse to shed new light on mammalian BM developmental dynamics.

Single Genomic Loci Labeling and Manipulation Using SIMBA System.

The SIMBA (Simultaneous Imaging and Manipulation of genomic loci by Biomolecular Assemblies) system is an innovative CRISPR-based imaging technique that leverages dCas9-SunTag and FRB-mCherry-HP1α, with scFv-FKBP acting as a bridge. This powerful system enables simultaneous visualization and manipulation of genomic loci. The dCas9-SunTag fusion protein allows for precise targeting of specific genomic sites, and the FRB-mCherry-HP1α fusion protein facilitates the condensation of chromatin at the targeted loci. The scFv-FKBP bridge protein links dCas9-SunTag and FRB-mCherry-HP1α, ensuring efficient and specific recruitment of HP1α to the desired genomic loci. This integrated approach allows us to visualize and manipulate genomic regions of interest, opening up new avenues for studying genome organization, gene expression regulation, and chromatin dynamics in living cells. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Cloning of genetic constructs Basic Protocol 2: Transient transfection in mammalian cells and live-cell imaging Basic Protocol 3: Generation of SIMBA-expressing stable cell lines Basic Protocol 4: Manipulation of genomic loci using SIMBA.

Role of Plasmonic Antenna in Hot Carrier-Driven Reactions on Bimetallic Nanostructures.

Noble metal nanostructures can efficiently harvest electromagnetic radiation, which, in turn, is used to generate localized surface plasmon resonances. Surface plasmons decay, producing hot carriers, that is, short-lived species that can trigger chemical reactions on metallic surfaces. However, noble metal nanostructures catalyze only a very small number of chemical reactions. This limitation can be overcome by coupling such nanostructures with catalytic-active metals. Although the role of such catalytically active metals in plasmon-driven catalysis is well-understood, the mechanistics of a noble metal antenna in such chemistry remains unclear. In this study, we utilize tip-enhanced Raman spectroscopy, an innovative nanoscale imaging technique, to investigate the rates and yields of plasmon-driven reactions on mono- and bimetallic gold- and silver-based nanostructures. We found that silver nanoplates (AgNPs) demonstrate a significantly higher yield of 4-nitrobenzenehtiol to p,p'-dimercaptoazobisbenzene (DMAB) reduction than gold nanoplates (AuNPs). We also observed substantially greater yields of DMAB on silver-platinum and silver-palladium nanoplates (Ag@PtNPs and Ag@PdNPs) compared to their gold analogues, Au@PtNPs and Au@PdNPs. Furthermore, Ag@PtNPs exhibited enhanced reactivity in 4-mercatophenylmethanol to 4-mercaptobenzoic acid oxidation compared to Au@PtNPs. These results showed that silver-based bimetallic nanostructures feature much greater reactivity compared to their gold-based analogues.

Circulating Cardiovascular Biomarkers in Cancer Therapeutics-Related Cardiotoxicity: Review of Critical Challenges, Solutions, and Future Directions.

Cardiotoxicity is a growing concern in the oncology population. Transthoracic echocardiography and multigated acquisition scans have been used for surveillance but are relatively insensitive and resource intensive. Innovative imaging techniques are constrained by cost and availability. More sensitive, cost-effective cardiotoxicity surveillance strategies are needed. Circulating cardiovascular biomarkers could provide a sensitive, low-cost solution. Biomarkers such as troponins, natriuretic peptides (NPs), novel upstream signals of oxidative stress, inflammation, and fibrosis as well as panomic technologies have shown substantial promise, and guidelines recommend baseline measurement of troponins and NPs in all patients receiving potential cardiotoxins. Nonetheless, supporting evidence has been hampered by several limitations. Previous reviews have provided valuable perspectives on biomarkers in cancer populations, but important analytic aspects remain to be examined in depth. This review provides comprehensive assessment of critical challenges and solutions in this field, with focus on analytical issues relating to biomarker measurement and interpretation. Examination of evidence pertaining to common and serious forms of cardiotoxicity reveals that improved study designs incorporating larger, more diverse populations, registry-based approaches, and refinement of current definitions are key. Further efforts to harmonize biomarker methodologies including centralized biobanking and analyses, novel decision limits, and head-to-head comparisons are needed. Multimarker algorithms incorporating machine learning may allow rapid, personalized risk assessment. These improvements will not only augment the predictive value of circulating biomarkers in cardiotoxicity but may elucidate both direct and indirect relationships between cardiovascular disease and cancer, allowing biomarkers a greater role in the development and success of novel anticancer therapies.

Innovative Imaging Techniques: A Conceptual Exploration of Multi-Modal Raman Light Sheet Microscopy.

Advances in imaging of microscopic structures are supported and complemented by adaptive visualization tools. These tools enable researchers to precisely capture and analyze complex three-dimensional structures of different kinds such as crystals, microchannels and electronic or biological material. In this contribution, we focus on 3D cell cultures. The new possibilities can play a particularly important role in biomedical research, especially here in the study of 3D cell cultures such as spheroids in the field of histology. By applying advanced imaging techniques, detailed information about the spatial arrangement and interactions between cells can be obtained. These insights help to gain a better understanding of cellular organization and function and have potential implications for the development of new therapies and drugs. In this context, this study presents a multi-modal light sheet microscope designed for the detection of elastic and inelastic light scattering, particularly Rayleigh scattering as well as the Stokes Raman effect and fluorescence for imaging purposes. By combining multiple modalities and stitching their individual results, three-dimensional objects are created combining complementary information for greater insight into spatial and molecular information. The individual components of the microscope are specifically selected to this end. Both Rayleigh and Stokes Raman scattering are inherent molecule properties and accordingly facilitate marker-free imaging. Consequently, altering influences on the sample by external factors are minimized. Furthermore, this article will give an outlook on possible future applications of the prototype microscope.

The role of innovative modeling and imaging techniques in improving outcomes in patients with LVAD.

Heart failure remains a significant cause of mortality in the United States and around the world. While organ transplantation is acknowledged as the gold standard treatment for end stage heart failure, supply is limited, and many patients are treated with left ventricular assist devices (LVADs). LVADs extend and improve patients' lives, but they are not without their own complications, particularly the hemocompatibility related adverse events (HRAE) including stroke, bleeding and pump thrombosis. Mainstream imaging techniques currently in use to assess appropriate device function and troubleshoot complications, such as echocardiography and cardiac computed tomography, provide some insight but do not provide a holistic understanding of pump induced flow alterations that leads to HRAEs. In contrast, there are technologies restricted to the benchtop-such as computational fluid dynamics and mock circulatory loops paired with methods like particle image velocimetry-that can assess flow metrics but have not been optimized for clinical care. In this review, we outline the potential role and current limitations of converging available technologies to produce novel imaging techniques, and the potential utility in evaluating hemodynamic flow to determine whether LVAD patients may be at higher risk of HRAEs. This addition to diagnostic and monitoring capabilities could improve prevention and treatment of LVAD-induced complications in heart failure patients.

Effect of the local energy distribution of x-ray beams generated through inverse Compton scattering in dual-energy imaging applications.

X-ray sources based on the inverse Compton interaction between a laser and a relativistic electron beam are emerging as a promising compact alternative to synchrotron for the production of intense monochromatic and tunable radiation. The emission characteristics enable several innovative imaging techniques, including dual-energy K-edge subtraction (KES) imaging. The performance of these techniques is optimal in the case of perfectly monochromatic x-ray beams, and the implementation of KES was proven to be very effective with synchrotron radiation. Nonetheless, the features of inverse Compton scattering (ICS) sources make them good candidates for a more compact implementation of KES techniques. The energy and intensity distribution of the emitted radiation is related to the emission direction, which means different beam qualities in different spatial positions. In fact, as the polar angle increases, the average energy decreases, while the local energy bandwidth increases and the emission intensity decreases. The scope of this work is to describe the impact of the local energy distribution variations on KES imaging performance. By means of analytical simulations, the reconstructed signal, signal-to-noise ratio, and background contamination were evaluated as a function of the position of each detector pixel. The results show that KES imaging is possible with ICS x-ray beams, even if the image quality slightly degrades at the detector borders for a fixed collimation angle and, in general, as the beam divergence increases. Finally, an approach for the optimization of specific imaging tasks is proposed by considering the characteristics of a given source.

Coronary sinus cannulation as real time anatomical landmark to facilitate left bundle branch area pacing

Abstract Funding Acknowledgements Type of funding sources: None. Background/Introduction Left bundle branch area pacing (LBBAP) is an established form of conduction system pacing. Innovative fluoroscopic imaging technique with use of radio-opaque contrast in the right ventricle or using pre-mapped His potential has been advocated as an acquired road map to assist in implantation of LBBAP(1) lead. Purpose We aim to explore a different technique by cannulating the coronary sinus (CS) with a sheath and wire to provide real-time anatomical guidance of location of the base of the heart to facilitate more effective LBBAP in a consistent fashion. Methods Prior to LBBAP, the body of the CS is cannulated using an AL2 diagnostic angiographic catheter and a 0.035-inch glidewire. The wire is fixated externally, and fluoroscopic image is acquired before proceeding with deployment of the pacing lead onto an optimal ventricular septal position via standard right (RAO 30) and left anterior oblique (LAO 30) views. From August 2021 to October 2022, we enrolled 12 consecutive patients requiring pacemaker for standard indications and explored the CS cannulation method in delivering LBBAP with either steerable catheters or fixed shape sheaths. Thereafter, we analysed and described the success rates, immediate electrical and device parameters, procedural durations, and clinical events. Results 12 patients were studied. The mean age was 75 ± 13.07 years and 6 (50%) patients were male. 1 (8.3%) was for sinus nodal dysfunction, 10 (83.3%) for atrioventricular block and 1 (8.3%) for atrial fibrillation (AF) control with atrioventricular node ablation. Mean left ventricular ejection fraction was 54.17 ± 8.21 % and baseline QRS was 108.08 ± 23.20ms. LBBAP was successful in all 12 patients based on published criteria with a mean paced QRS duration of 115.25 ± 9.28 ms. The mean total procedure duration was 130.75 +/- 29.84 minutes and mean fluoroscopic duration was 23.17 ± 9.11 minutes. After 1st week post implant, the pacing threshold, sensing, and impedances were stable and acceptable in all the patients. There were no acute complications (e.g. pneumothorax, lead complications, lead dislodgements / perforations) during the periprocedural period. Conclusion(s) The CS cannulation method is a feasible, safe, and effective method to to provide real-time anatomical visualisation of the base of the heart. This facilitates the achievement for optimal LBBAP and can be adopted especially in difficult cardiac anatomies or rotated cardiac silhouettes identified on fluoroscopy.

Less-invasive autopsy for early pregnancy loss.

Autopsy investigations provide valuable information regarding fetal death that can assist in the parental bereavement process, and influence future pregnancies, but conventional autopsy is often declined by parents because of its invasive approach. This has led to the development of less-invasive autopsy investigations based on imaging technology to provide a more accessible and acceptable choice for parents when investigating their loss. Whilst the development and use of more conventional clinical imaging techniques (radiographs, CT, MRI, US) are well described in the literature for fetuses over 20weeks of gestational age, these investigations have limited diagnostic accuracy in imaging smaller fetuses. Techniques such as ultra-high-field MRI (>3T) and micro-focus computed tomography have been shown to have higher diagnostic accuracy whilst still being acceptable to parents. By further developing and increasing the availability of these more innovative imaging techniques, parents will be provided with a greater choice of acceptable options to investigate their loss, which may in turn increase their uptake. We provide a narrative review focussing on the development of high-resolution, non-invasive imaging techniques to evaluate early gestational pregnancy loss.

Transmission-Based Muography for Ore Bodies Prospecting: A Case Study from a Skarn Complex in Italy

Transmission-based muography (TM) is an innovative imaging technique based on the measurement and analysis of the cosmic ray muons flux attenuation within the target under investigation. This technique allows imaging inner-body density differences and has successfully been applied in a wide range of research fields: geology, archaeology, engineering geology and civil engineering. The aim of this study is to show the reliability of TM as an innovative, noninvasive geophysical method for ore body prospecting and other mining related studies. The measurements were carried out at the Temperino mine in the San Silvestro Archaeological and Mining Park (Campiglia Marittima, Italy), where several magmatic and metasomatic geological units are embodied. Among them, a Cu–Fe–Zn–Pb(–Ag) sulfide skarn complex primarily composed by hedenbergite and ilvaite minerals. Using the acquired muon imaging data obtained with the MIMA (Muon Imaging for Mining and Archaeology) detector prototype (cubic detector of 0.5 × 0.5 × 0.5 m3), the presence of a high-density vein inside the skarn body within the rock volume above the muon detector was identified, localized and interpreted. Applying a back-projection algorithm to the obtained 2D transmission map made it possible to estimate and visualize as point cloud data, in a 2D or 3D environment, the identified high-density body and its relative distance from the detector. The results of this study highlight the potential of muography as a support tool to other geophysical methods in the field of mining exploration.

Multi-Technique Approach by Traditional and Innovative Methodologies to Support the Restoration of a Wall Painting from the 16th Century at Palazzo Gallo in Bagnaia, Viterbo, Central Italy

This paper summarizes the main results of the study, diagnostics, and restorations conducted and applied to a 16th century wall painting (a portion of the frieze and the Riario coat of arm) in Palazzo Gallo (Bagnaia, Viterbo district—central Italy) recently concluded, which was also the subject of a master’s degree thesis in Conservation and Restoration of Cultural Heritage at the University of Tuscia, Viterbo. Innovative imaging techniques were used for the first time on a wall painting on-site: hypercolorimetric multispectral imaging (HMI) and pulse-compression thermography (PuCT), combined with more traditional analysis such as X-ray fluorescence spectroscopy, Fourier transform infrared spectroscopy, and cross-section investigation. HMI allowed for mapping the conservation status before and after the removal of the scialbo layer that covered the original paintings. It also allowed different areas of the painting to be compared and for verifying the effectiveness of the cleaning. PuCT enabled the detection of cracks and discontinuities in the ground layers and to evaluate the depth of such anomalies, giving valuable support in the consolidation step. Moreover, passive thermography was used to monitor the penetration level of a hydraulic mortar in real time, a technique that was greatly helpful for verifying the successful fill and consolidation of voids beneath the pictorial layer. Overall, the multi-technique approach reported here was of considerable assistance for restoration of the mentioned artwork, the result of which has also been documented.

Nanoscale imaging of individual amyloid aggregates extracted from brains of Alzheimer and Parkinson patients reveals presence of lipids in α-synuclein but not in amyloid β1-42 fibrils.

Abrupt aggregation of misfolded proteins is the underlying molecular cause of Alzheimer disease (AD) and Parkinson disease (PD). Both AD and PD are severe pathologies that affect millions of people around the world. A small 42 amino acid long peptide, known as amyloid β (Aβ), aggregates in the frontal cortex of AD patients forming oligomers and fibrils, highly toxic protein aggregates that cause progressive neuron death. Similar aggregates of α-synuclein (α-Syn), a small protein that facilitates neurotransmitter release, are observed in the midbrain, hypothalamus, and thalamus of people with PD. In this study, we utilized the innovative nano-Infrared imaging technique to investigate the structural organization of individual Aβ and α-syn fibrils postmortem extracted from brains of AD and PD patients, respectively. We observed two morphologically different Aβ and α-Syn fibril polymorphs in each patient's brain. One had twisted topology, whereas another exhibited flat tape-like morphology. We found that both polymorphs shared the same parallel β-sheet-dominated secondary structure. These findings suggested that both fibril polymorphs were built from structurally similar if not identical filaments that coiled forming twisted fibrils or associated side-by-side in the case of straight Aβ and α-Syn fibrils. Nano-Infrared analysis of individual protein aggregates also revealed the presence of lipids in the structure of both twisted and tape-like α-Syn fibrils that were not observed in any of the Aβ fibril polymorphs. These findings demonstrate that lipid membranes can play a critically important role in the onset and progression of PD.

Ocular alterations in patients with Alport syndrome-An update

Alport syndrome (AS) is arare hereditary systemic disease that results in alterations of the kidneys, inner ear, and various structures of the eye. It is caused by mutations in one of the genes encoding collagen typeIV. In recent years, new and innovative imaging techniques have added characteristics of ocular alterations in AS and provided new insights, including into the pathogenesis of the disease. The aim of this paper is to provide an overview of the current knowledge of ocular changes in AS, as well as to present the Alport ocular pass. Narrative review article. Ocular manifestations of AS include changes in the cornea, lens, and retina. Specifically, posterior polymorphic corneal dystrophy, anterior lenticonus (pathognomonic for AS), and various retinal changes have been described, which have been further characterized in recent years by newer imaging techniques. In particular, foveal changes in AS may present as both athickened central retina in the context of foveal hypoplasia or astaircase-like thinning of the fovea. Both lesions could provide further insights into the role of typeIV collagen in ocular structures. The AS can manifest in various structures of the eye. The staircase-like changes of the central retina in AS patients indicate the important role of collagen typeIV in the homeostasis and regular function of the inner retinal layers. The often mild foveal hypoplasia may provide clues to the role of collagen typeIV in retinal embryogenesis. While anterior lenticonus is pathognomonic for AS and can be treated easily by refractive lens exchange, the only option currently available for retinal alterations is close follow-up and, if necessary, treatment of systemic complications of AS.

Non-invasive imaging techniques in presurgical margin assessment of basal cell carcinoma: Current evidence.

The diagnosis of basal cell carcinoma (BCC) is based on clinical and dermoscopical features. In uncertain cases, innovative imaging techniques, such as reflectance confocal microscopy (RCM) and optical coherence tomography (OCT), have been used. The main limitation of these techniques is the inability to study deep margins. HFUS (high-frequency ultrasound) and the most recent UHFUS (ultra-high-frequency ultrasound) have been used in various applications in dermatology, but they are not yet routinely used in the diagnosis of BCC. A key point in clinical practice is to find an imaging technique that can help to reduce post-surgical recurrences with a careful presurgical assessment of the lesional margins. This technique should show high sensitivity, specificity, reproducibility and simplicity of execution. This concept is very important for the optimal management of patients who are often elderly and have many comorbidities. The aim of the paper is to analyse the characteristics of current imaging techniques and the studies in the literature on this topic. The authors independently searched the MEDLINE, PubMed, Embase, Scopus, ScienceDirect and Cochrane Library databases for studies looking for non-invasive imaging techniques for the presurgical margin assessment of BCC. Preoperative study of the BCC subtype can help to obtain a complete excision with free margins. Different non-invasive imaging techniques have been studied for in vivo evaluation of tumour margins, comparing the histologic evaluation with a radical surgery. The possibility to study the lateral and deep margins would allow a reduction of recurrences and sparing of healthy tissue. HFUS and UHFUS represent the most promising, non-invasive techniques for the pre-operative study of BCC facilitating the characterization of vascularization, deep lateral margins and high-risk subtypes, although they are limited by insufficient literature unlike RCM and OCT.

Super-resolution 3D maps to study the antisecretory factor's effects on different GABAa receptor subunits in rat cerebellar granule cells in vitro.

The Antisecretory Factor (AF) is an endogenous protein that inhibits intestinal hypersecretion and various inflammation disorders in vivo. AF has been detected in many mammalian tissues and plasma, but its mechanisms of action are essentially unknown. Previously, we studied the pharmacological action of the AF on GABAA receptors in cerebellar granule cells, comparing the electrophysiological response evoked by two-photon mediated release of caged GABA compounds before and after the administration of AF-16, a 16 amino acids long peptide obtained from the amino-terminal end of the AF protein. After the treatment with AF-16, we observed an increase in the GABAA receptor responses, particularly in those containing the α6 subunit. To figure out the interactions of AF with GABAA receptors in the same cellular model (cerebellar granule cells), we combined immunofluorescence subunits’ staining with confocal and super-resolution microscopy. In particular, we took advantage of an innovative imaging technique that combines stimulated emission depletion (STED) with fluorescence lifetime microscopy (FLIM) to collect super-resolution 3D maps of different subunits distributed on the neuron cell membrane. We explored different approaches to analyze super-resolution fluorescence images obtained by labeling α1 and α6 subunits before and after 1-hour incubation with AF-16. Comparing pre- and post-treatment maps, we found differences in how different subunit populations respond to AF treatment. We critically analyzed these new experimental findings with our previous electrophysiological data to widen the knowledge of the mechanisms of interaction between GABAA receptor subunits and AF protein.