A5010539283- Cell Image Analysis Techniques
- Cardiac electrophysiology and arrhythmias
- Advanced Fluorescence Microscopy Techniques
- Neuroscience and Neural Engineering
- Photoreceptor and optogenetics research
- Neural dynamics and brain function
- Anatomy and Medical Technology
- Single-cell and spatial transcriptomics
- Ion channel regulation and function
- Cardiomyopathy and Myosin Studies
- Optical Imaging and Spectroscopy Techniques
- Optical Coherence Tomography Applications
- bioluminescence and chemiluminescence research
- Cardiovascular Function and Risk Factors
- Digital Holography and Microscopy
- Advanced Neuroimaging Techniques and Applications
- Functional Brain Connectivity Studies
- Cardiovascular Effects of Exercise
- Cardiac Arrhythmias and Treatments
- Anesthesia and Pain Management
- Pediatric Pain Management Techniques
- Medical Image Segmentation Techniques
- ATP Synthase and ATPases Research
- Electrospun Nanofibers in Biomedical Applications
- Cellular transport and secretion
University of Florence
2016-2025
European Theoretical Spectroscopy Facility
2024
Centre Hospitalier de Lens
2021
National Research Council
2017-2020
National Institute of Optics
2018
WWF Italia
2015-2017
King's College London
2016
Centro Cardiologico Monzino
2016
Institute of Experimental Cardiology
2016
Imperial College London
2016
Abstract Current rescue therapies for life-threatening arrhythmias ignore the pathological electro-anatomical substrate and base their efficacy on a generalized electrical discharge. Here, we developed an all-optical platform to examine less invasive defibrillation strategies. An ultrafast wide-field macroscope was optically map action potential propagation with red-shifted voltage sensitive dye in whole mouse hearts. The implemented random-access scanning head capable of drawing...
Disopyramide is effective and safe in patients with obstructive hypertrophic cardiomyopathy. However, its cellular molecular mechanisms of action are unknown. We tested disopyramide cardiomyocytes from the septum surgical myectomy patients: inhibits multiple ion channels, leading to lower Ca transients force, shortens potentials, thus reducing arrhythmias. The electrophysiological profile explains efficient reduction outflow gradients but also limited prolongation QT interval absence...
The combination of optical tissue transparency with immunofluorescence allows the molecular characterization biological tissues in 3D. However, adult human organs are particularly challenging to become transparent because autofluorescence contributions aged tissues. To meet this challenge, we optimized SHORT (SWITCH-H
Brain cells are arranged in laminar, nuclear, or columnar structures, spanning a range of scales. Here, we construct reliable cell census the frontal lobe human cerebral cortex at micrometer resolution magnetic resonance imaging (MRI)-referenced system using innovative and analysis methodologies. MRI establishes macroscopic reference coordinate laminar cytoarchitectural boundaries. Cell counting is obtained with digital stereological approach on 3D reconstruction cellular from custom-made...
Abnormalities of cardiomyocyte Ca2+ homeostasis and excitation–contraction (E–C) coupling are early events in the pathogenesis hypertrophic cardiomyopathy (HCM) concomitant determinants diastolic dysfunction arrhythmias typical disease. T-tubule remodelling has been reported to occur HCM but little is known about its role E–C alterations HCM. Here, electro-mechanical associated investigated Δ160E cTnT mouse model that expresses a clinically-relevant mutation. Contractile function intact...
Although optogenetics has clearly demonstrated the feasibility of cardiac manipulation, current optical stimulation strategies lack capability to react acutely ongoing wave dynamics. Here, we developed an all-optical platform monitor and control electrical activity in real-time. The methodology was applied restore normal after atrioventricular block manipulate intraventricular propagation wavefront. closed-loop approach also simulate a re-entrant circuit across ventricle. development this...
Rationale: Recently, abundant axial tubule (AT) membrane structures were identified deep inside atrial myocytes (AMs). Upon excitation, ATs rapidly activate intracellular Ca2+ release and sarcomeric contraction through extensive AT junctions, a cell-specific mechanism. While junctions with the sarcoplasmic reticulum contain unusually large clusters of ryanodine receptor 2 (RyR2) channels in mouse AMs, it remains unclear if similar protein networks exist across species, particularly those...
Significance The homogenous propagation of the action potential in cardiac cells is guaranteed by a complex network membrane invaginations called T-tubular system. In diseases, T-tubules may show electrical defects that can compromise cell function. Here, we investigate diffusional properties fluorescent probes inside to predict conductivity tubular network. We apply this method detecting alterations T-tubule pathological setting characterized compromised integrity. found heart failure,...
Rationale: The sarcolemma of cardiomyocytes contains many proteins that are essential for electromechanical function in general, and excitation-contraction coupling particular. distribution these is nonuniform between the bulk sarcolemmal surface membrane invaginations known as transverse tubules (TT). TT form an intricate network fluid-filled conduits support synchronicity within cardiomyocytes. Although continuous with extracellular space, narrow lumen tortuous structure can domains...
Obscurin is a large muscle protein whose multiple functions include providing mechanical strength to the M-band and linking sarcomere sarcoplasmic reticulum. Mutations in obscurin are linked various forms of diseases. This study compares cardiac function murine model deletion (KO) with wild-type (WT) vivo ex vivo. Echocardiography showed that KO hearts had larger (+20%) end-diastolic end-systolic volumes, reduced fractional shortening, impaired ejection fraction, consistent dilated...
3D reconstruction of human brain volumes at high resolution is now possible thanks to advancements in tissue clearing methods and fluorescence microscopy techniques. Analyzing the massive data produced with these approaches requires automatic able perform fast accurate cell counting localization. Recent advances deep learning have enabled development various tools for segmentation. However, quantification neurons presents specific challenges, such as pixel intensity variability,...
Accurate labeling of specific layers in the human cerebral cortex is crucial for advancing our understanding neurodevelopmental and neurodegenerative disorders. Building on recent advancements ultra-high-resolution ex vivo MRI, we present a novel semi-supervised segmentation model capable identifying supragranular infragranular MRI with unprecedented precision. On dataset consisting 17 whole-hemisphere scans at 120 $\mu $m, propose Multi-resolution U-Nets framework that integrates global...
ABSTRACT This study explores the use of polarized second-harmonic generation (pSHG) to investigate myosin conformation in relaxed state, differentiating between actin-available, disordered (ON) state and energy-conserving, ordered (OFF) state. By pharmacologically shifting ON/OFF equilibrium using a activator (2-deoxyATP) or inhibitor (Mavacamten), we demonstrate sensitivity pSHG quantifying ratio skeletal cardiac tissues. Comparisons with X-ray diffraction measurements further validate our...
The combination of tissue clearing techniques with advanced optical microscopy facilitates the achievement three-dimensional (3D) reconstruction macroscopic specimens at high resolution. Whole mouse organs or even bodies have been analyzed, while human nervous system remains a challenge. Although several protocols proposed, autofluorescence and variable post-mortem conditions negatively affect quality images in terms achievable transparency staining contrast. Moreover, homogeneous...
Abstract T‐tubules (TT) form a complex network of sarcolemmal membrane invaginations, essential for well‐co‐ordinated excitation–contraction coupling (ECC) and thus homogeneous mechanical activation cardiomyocytes. ECC is initiated by rapid depolarization the membrane. Whether TT active (local generation action potentials; AP) or passive (following outer cell surface sarcolemma; SS) has not been experimentally validated in Based on assessment ion flux pathways needed AP generation, we...
Axial tubule junctions with the sarcoplasmic reticulum control rapid intracellular Ca2+-induced Ca2+ release that initiates atrial contraction. In myocytes we previously identified a constitutively increased ryanodine receptor (RyR2) phosphorylation at junctional sites, whereas non-junctional RyR2 clusters were phosphorylated acutely following β-adrenergic stimulation. Here, hypothesized baseline synthesis of 3',5'-cyclic adenosine monophosphate (cAMP) is augmented in axial compartments...
Accurate labeling of specific layers in the human cerebral cortex is crucial for advancing our understanding neurodevelopmental and neurodegenerative disorders. Leveraging recent advancements ultra-high resolution ex vivo MRI, we present a novel semi-supervised segmentation model capable identifying supragranular infragranular MRI with unprecedented precision. On dataset consisting 17 whole-hemisphere scans at 120 μm, propose multi-resolution U-Nets framework (MUS) that integrates global...
Despite the numerous clearing techniques that emerged in last decade, processing postmortem human brains remains a challenging task due to its dimensions and complexity, which make imaging with micrometer resolution particularly difficult. This paper presents protocol perform reconstruction of volumetric portions brain by simultaneously tens sections SHORT (SWITCH - H2O2 Antigen Retrieval 2,2'-thiodiethanol [TDE]) tissue transformation protocol, enables clearing, labeling, sequential samples...
Asynchronous Ca2+ release promotes non-homogeneous myofilament activation, leading to mechanical dysfunction, as well initiation of propagated calcium waves and arrhythmias. Recent advances in microscopy techniques allowed functional measurements cardiac preparation with unprecedented spatial temporal resolution. Ultra-fast deflectors were used rapidly scan laser beams across the sample, performing optical local fluxes action potential from multiple sites within cells. The capability probing...
High-speed imaging with light-sheet fluorescence microscopy poses several challenges throughout the whole pipeline, from data acquisition in lab to image stitching and post-processing. Here we present our custom hardware software solutions that allow us map large biological samples at cellular level, e.g. portions of human brain cortex. Our optical setup—a dual-view, inverted, microscope—is capable simultaneous two-color a rate 1 GB/s. open source tools include instrument's control also...
Optogenetics is an emerging method that uses light to manipulate electrical activity in excitable cells exploiting the interaction between and light-sensitive depolarizing ion channels, such as channelrhodopsin-2 (ChR2). Initially used neuroscience, it has been adopted cardiac research where expression of ChR2 preparations allows optical pacing, resynchronization defibrillation. Recently, optogenetics leveraged intact heart real-time. This new approach was applied simulate a re-entrant...