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Faccio, D.

Publications and source records attributed to Faccio, D..

4 recordsLinked to original sources

Fluorescence-lifetime optical electrophysiology in contracting cardiomyocytes

Precise monitoring of cardiac electrophysiology in vitro is crucial to understanding heart function and cardiac disease. However, high-throughput, contact-free methods for directly measuring excitation-contraction coupling remain limited. Here, we introduce a new paradigm for quantitative electrophysiological imaging that combines fluorescence lifetime and intensity information to capture dynamic cardiac signals with high fidelity. We show that lifetime measurements are intrinsically decoupled from motion artifacts and provide calibrated calcium concentration and membrane potential readouts across wide fields of view. Using a gated single-photon avalanche diode camera, we acquire fluorescence lifetime images at up to 200 frames per second with sufficient signal-to-noise ratio such that each frame contains meaningful lifetime information without temporal averaging. This approach yields spatially resolved maps of absolute voltage and calcium values across contracting cardiomyocyte monolayers, revealing heterogeneous cell behaviors within individual assays and uncovering previously unreported dynamics during late-phase repolarization for real-time analysis of excitation-contraction coupling.

bioengineering↗

Real-Time Wide-Field Fluorescence Lifetime Imaging via Single-Snapshot Acquisition for Biomedical Applications

Fluorescence lifetime imaging (FLI) is a powerful tool for investigating molecular processes, microenvironmental parameters, and molecular interactions across tissue to (sub-)cellular levels. Despite its established value in numerous biomedical applications, conventional FLI techniques are hindered by long acquisition times. This limitation restricts their use in real-time scenarios, such as monitoring fast biological processes, studying live organisms, and in environments that require rapid imaging and immediate inference, such as clinical image-guided interventions. Here, we present a novel FLI approach that combines a large-format time-gated SPAD array with dual-gate acquisition capability, alongside a rapid lifetime determination algorithm. This integration allows for real-time fluorescence lifetime estimation through single-snapshot acquisitions, eliminating the need for traditional, time-consuming time-resolved data collection. We demonstrate the scalability and versatility of this method by achieving real-time FLI across challenging biomedical applications, ranging from capturing fast neural dynamics at the microscopic scale, performing multimodal 3D volumetric FLI of tumor organoids at the mesoscopic scale, to macroscale FLI in both direct and highly scattering regimes. Furthermore, we validate its utility in fluorescence lifetime-guided surgical procedures using tissue-mimicking phantoms. Overall, this new methodology significantly enhances the temporal and spatial capabilities of FLI, opening the door to the assessment of fast dynamic biomedical signals. It also enables the seamless integration of FLI into clinical workflows, particularly in applications like fluorescence-guided surgery.

bioengineering↗

Wide-field Fluorescence Lifetime Imaging Microscopy with a High-Speed Mega-pixel SPAD Camera

Fluorescence lifetime imaging microscopy (FLIM) is a key technology that provides direct insight into cell metabolism, cell dynamics and protein activity. However, determining the lifetimes of different fluorescent proteins requires the detection of a relatively large number of photons, hence slowing down total acquisition times. Moreover, there are many cases, for example in studies of cell collectives, where wide-field imaging is desired. We report scan-less wide-field FLIM based on a 0.5 Megapixel resolution, time-gated Single Photon Avalanche Diode (SPAD) camera, with acquisition rates up to 1 Hz. Fluorescence lifetime estimation is performed via a pre-trained artificial neural network with 1000-fold improvement in processing times compared to standard least squares fitting techniques. We utilised our system to image HT1080 - human fibrosarcoma cell line as well as Convallaria. The results show promise for real-time FLIM and a viable route towards multi-megapixel fluorescence lifetime images, with a proof-of-principle mosaic image shown with 3.6 megapixels.

bioengineering↗

Multifunction control and evaluation of a 3D printed hand prosthesis with the Myo armband by hand amputees

Upper limb amputations are highly impairing injuries that can substantially limit the quality of life of a person. The most advanced dexterous prosthetic hands have remarkable mechanical features. However, in most cases, the control systems are a simple extension of basic control protocols, making the use of the prosthesis not intuitive and sometimes complex. Furthermore, the cost of dexterous prosthetic hands is often prohibitive, especially for the pediatric population and developing countries. 3D printed hand prostheses can represent an opportunity for the future. Open 3D models are increasingly being released, even for dexterous prostheses that are capable of moving each finger individually and actively rotating the thumb. However, the usage and test of such devices by hand amputees (using electromyography and classification methods) is not well explored. The aim of this article is to investigate the usage of a cost-effective system composed of a 3D printed hand prosthesis and a low-cost myoelectric armband. Two subjects with transradial amputation were asked to wear a custom-made socket supporting the HANDi Hand and the Thalmic Labs Myo armband. Afterwards, the subjects were asked to control and use the prosthetic hand to grasp several objects by attempting to perform a set of different hand gestures. Both the HANDi Hand and the Myo armband performed well during the test, which is encouraging considering that the HANDi Hand was developed as a research platform. The results are promising and show the feasibility of the multifunction control of dexterous 3D printed hand prostheses based on low-cost setups. Factors as the level of the amputation, neuromuscular fatigue and mechanical limitations of the 3D printed hand prosthesis can influence the performance of the setup. Practical aspects such as usability and robustness will need to be addressed for successful application in daily life. A video of the tests can be found at the following link: https://youtu.be/iPSCAbd17Qw

bioengineering↗