Biology subjects

Meneghetti, M.

Publications and source records attributed to Meneghetti, M..

10 recordsLinked to original sources

bioRxiv2025

Multi-spectral optoacoustic microscopy driven by gas-filled hollow-core fiber laser pulses

Multi-spectral optoacoustic microscopy (MS-OAM) requires high-performance light sources capable of delivering multiple intense spectral lines precisely matched to the absorption characteristics of selected biomolecules. We present a gas-filled anti-resonant hollow-core fiber (ARHCF) laser source optimized for near-infrared (NIR) MS-OAM. The hydrogen (H2)-filled ARHCF laser emits multiple spectral lines with high pulse energy and narrow linewidths (<0.1 nm) across a broad spectral range ([~]1100 nm to [~]2200 nm). Several Raman laser lines were generated to overlap with key biomolecular absorption bands, including lipids (1210 nm and 1700 nm), collagen ([~]1540 nm), and water ([~]1400 nm and [~]1900 nm). We demonstrate the systems performance by mapping absorbers in the first and second overtone regions of hair, pig tissue, and collagen samples. This work aims to bring the gas-filled fiber technology in MS-OAM applications and paves the way for high-resolution, label-free bio-imaging across extended infrared and ultraviolet regimes.

biophysics↗

bioRxiv2024

Soft biocompatible polymer optical fiber tapers for implantable neural devices

Optical fibers are between the most common implantable devices for delivering light in the nervous system for optogenetics and infrared neural stimulation applications. Tapered optical fibers, in particular, can offer homogeneous light delivery to a large volume and spatially resolved illumination compared to standard flat-cleaved fibers while being minimally invasive. However, the use of tapers for neural applications has up to now been limited to silica optical fibers, whose large Youngs modulus can cause detrimental foreign body response in chronic settings. Here, we present the fabrication and optimization of tapered fiber implants based on polymer optical fibers (POFs). After numerically determining the optimal materials and taper geometry, we fabricated two types of POFs by thermal fiber drawing. The fabrication of the taper was achieved by chemical etching of the fibers, for which several solvents previously reported in literature have been tested. The influence of different parameters on the etching process and on the quality of the obtained tapers was also investigated. The large illumination volume of the produced high-quality taper-based implants was finally tested in vitro in a brain phantom.

neuroscience↗

bioRxiv2024

Lifting the degeneracy of optical fiber neural interfaces for optogenetics and electrophysiology by using axialtrodes

Modulating and recording neuronal activity are essential for probing brain function and developing therapies for neurological disorders. However, conventional flat-end optical fibers--widely used for deep brain access-- interact with neural tissue only at their distal tip, limiting spatial resolution across brain layers. To overcome this constraint, we introduce the microfluidic axialtrode: a soft, multimodal neural interface created by controllably angle-cleaving a thermally drawn, multimaterial optical fiber integrated with multiple metal electrodes and microfluidic channels. We demonstrate in vivo that this design enables spatially distributed optogenetics, multisite electrophysiological recording, and targeted drug delivery along the fibers axis, allowing simultaneous interaction with multiple neuronal layers. The axial configuration increases the functional interface with brain tissue, while the soft polymer construction and reduced cross-sectional footprint significantly suppress the inflammatory response compared to conventional silica fibers. Integration with a 3D-printed scaffold, fabricated from FDA-approved biocompatible resin, provides mechanical stability and compatibility with standard experimental hardware. The monolithic integration of these features positions the axialtrode as a scalable and versatile platform for next-generation neural interfacing.

neuroscience↗

bioRxiv2024

In vivo evaluation of thermally drawn biodegradable optical fibers as brain neural interfaces

Optical fiber technology has a critical role in modern neuroscience towards understanding the complex neuronal dynamics within the nervous system. In this study, we manufactured amorphous thermally drawn poly D, L-lactic acid (PDLLA) optical fibers in different diameters. These fibers were then implanted into the lateral posterior (LP) region of the mouse brain for 4 months, allowing us to assess their degradation characteristics. The gradual dissolution of the implanted PDLLA optical fibers in the brain was confirmed by optical microscope and scanning electron microscopy (SEM), molecular weight measurements, and light transmission spectroscopy. The results indicate that the degradation rate was mainly pronounced during the first week. Degradation after 4 months resulted in the formation of micropores on the surface of the implanted fiber within the gray matter region of the brain. Moreover, the current PDLLA optical fiber implant offers efficient light transmission in the short-wavelength near-infrared (SW-NIR) range. SW-NIR enables deeper tissue penetration and reduced light scattering, making it ideal for optogenetics and functional imaging with therapeutic potential in neurological disorders. We believe that the provided PDLLA optical fiber in this study constitutes a promising candidate for the development of next-generation biocompatible, soft, and biodegradable bi-directional neural interfaces

bioengineering↗

bioRxiv2024

Mid-infrared photoacoustic brain imaging enabled by cascaded gas-filled hollow-core fiber lasers

SignificanceExtending the photoacoustic microscopy (PAM) into the mid-infrared (MIR) molecular fingerprint region constitutes a promising route towards label-free imaging of biological molecular structures. Realizing this objective requires a high-energy nano-second MIR laser source. However, existing MIR laser technologies are limited to either low pulse energy or free-space structure which is sensitive to environmental conditions. Fiber lasers are promising technologies for PAM for their potential of offering both high pulse energy and robust performance against environmental conditions. However, MIR high energy fiber laser has not yet been used for PAM because it is still at the infant research stage. AimWe aim to employ the emerging gas-filled anti-resonant hollow-core fiber (ARHCF) laser technology for MIR-PAM for the purpose of imaging myelin-rich regions in a mouse brain. ApproachThis laser source is developed with a [~]2.75 J high-pulse-energy nano-second laser at 3.4 m, targeting the main absorption band of myelin sheaths, the primary chemical component of axons in the central nervous system. The laser mechanism relies on two-orders gas-induced vibrational stimulated Raman scattering (SRS) for nonlinear wavelength conversion, starting from a 1060 nm pump laser to 1409 nm through the 1st order Stokes generation in the nitrogen-filled 1st stage ARHCF, then, from 1409 nm to 3.4 m through the 2nd stage hydrogen-filled ARHCF. ResultsThe developed Raman laser was used for the first time for transmission-mode MIR-PAM of mouse brain regions containing rich myelin structures. ConclusionsThis work pioneers the potential use of high-energy and nano-second gas-filled ARHCF laser source to MIR-PAM, with a first attempt to report this kind of fiber laser source for PAM of lipid-rich myelin regions in a mouse brain. The proposed ARHCF laser technology is also expected to generate high-energy pulses at the ultraviolet (UV) region, which can significantly improve the lateral resolution of the PAM.

neuroscience↗

bioRxiv2023

In vivo multi-site electrophysiology enabled by flexible optrodes towards bi-directional spinal cord interrogation

Optical neural interfaces combining optogenetics and electrophysiology have been demonstrated as powerful tools for distinguishing the causal roles of neural circuits in the nervous system. Functional optrodes for multipoint stimulation and recording have already been demonstrated in the brain. However, soft and flexible multimodal optrodes for the purpose of probing the spinal cord have remained undeveloped. Here, we present the design and fabrication of a novel optrode for multi-site optical stimulation and electrical recording in the spinal cord by combining optical fiber drawing of polymer material, laser micromachining, and integration of tungsten microelectrodes in a monolithic fiber-based structure. The results from space-resolved scattering measurements, electrochemical impedance spectroscopy, and an acute in vivo electrophysiology experiment in an anesthetized rodent suggest this probe as a potential novel interface, which can serve as a part of therapeutic strategies against neurological conditions and injury in the spinal cord.

neuroscience↗

bioRxiv2022

Mapping whole brain effects of infrared neural stimulation with positron emission tomography

The combination of neuroimaging and targeted neuromodulation is a crucial tool to gain a deeper understanding of neural networks at a circuit level. Infrared neurostimulation (INS) is a promising optical modality that allows to evoke neuronal activity with high spatial resolution without need for the introduction of exogenous substances in the brain. Here, we report the use of whole-brain functional [18F]fluorodeoxyglucose positron emission tomography (FDG-PET) imaging during INS in the dorsal striatum, performed using a multifunctional soft neural probe. We demonstrate the possibility to identify multi-circuit connection patterns in both cortical and subcortical brain regions within a single scan. By using a bolus plus infusion FDG-PET scanning protocol, we were able to observe the metabolic rate evolution in these regions during the experiments and correlate its variation with the onset of the INS stimulus. Due to the focality of INS and the large amount of viable molecular targets for PET, this novel approach to simultaneous imaging and stimulation is highly versatile. This pilot study can pave the way to further understand the brain connectivity on a global scale.

neuroscience↗

bioRxiv2022

Opsin-free optical neuromodulation and electrophysiology enabled by a soft monolithic infrared multifunctional neural interface

Controlling neuronal activity with high spatial resolution using multifunctional and minimally invasive neural interfaces constitutes an important step towards developments in neuroscience and novel treatments for brain diseases. While infrared neuromodulation is an emerging technology for controlling the neuronal circuitry, it lacks soft implantable monolithic interfaces capable of simultaneously delivering light and recording electrical signals from the brain while being mechanically brain-compatible. Here, we have developed a soft fibre-based device based on high-performance thermoplastics which are >100-fold softer than silica glass. The presented fibre-implant is capable of safely neuromodulating the brain activity in localized cortical domains by delivering infrared laser pulses in the 2 m spectral region while recording electrophysiological signals. Action and local field potentials were recorded in vivo in adult rats while immunohistochemical analysis of the tissue indicated limited microglia and monocytes response introduced by the fibre and the infrared pulses. We expect our devices to further enhance infrared neuromodulation as a versatile approach for fundamental research and clinically translatable therapeutic interventions.

neuroscience↗

bioRxiv2020

Heparin inhibits cellular invasion by SARS-CoV-2: structural dependence of the interaction of the surface protein (spike) S1 receptor binding domain with heparin.

The dependence of the host on the interaction of hundreds of extracellular proteins with the cell surface glycosaminoglycan heparan sulphate (HS) for the regulation of homeostasis is exploited by many microbial pathogens as a means of adherence and invasion. The closely related polysaccharide heparin, the widely used anticoagulant drug, which is structurally similar to HS and is a common experimental proxy, can be expected to mimic the properties of HS. Heparin prevents infection by a range of viruses if added exogenously, including S-associated coronavirus strain HSR1. Heparin prevents infection by a range of viruses if added exogenously, including S-associated coronavirus strain HSR1. Here, we show that the addition of heparin to Vero cells between 6.25 - 200 g.ml-1, which spans the concentration of heparin in therapeutic use, and inhibits invasion by SARS-CoV-2 at between 44 and 80%. We also demonstrate that heparin binds to the Spike (S1) protein receptor binding domain and induces a conformational change, illustrated by surface plasmon resonance and circular dichroism spectroscopy studies. The structural features of heparin on which this interaction depends were investigated using a library of heparin derivatives and size-defined fragments. Binding is more strongly dependent on the presence of 2-O or 6-O sulphation, and the consequent conformational consequences in the heparin structure, than on N-sulphation. A hexasaccharide is required for conformational changes to be induced in the secondary structure that are comparable to those that arise from heparin binding. Enoxaparin, a low molecular weight clinical anticoagulant, also binds the S1 RBD protein and induces conformational change. These findings have implications for the rapid development of a first-line therapeutic by repurposing heparin as well as for next-generation, tailor-made, GAG-based antiviral agents against SARS-CoV-2 and other members of the Coronaviridae.

biochemistry↗

bioRxiv2020

Heparan sulfate structure is influenced by the ER-Golgi dynamics of its modifying enzymes

The cell surface and extracellular matrix polysaccharide, heparan sulfate (HS) conveys chemical information to control or influence crucial biological processes. Attempts to describe its structure-function relationships with HS binding proteins in a classical lock and key type manner, however, have been unsuccessful. HS chains are synthesized in a non-template driven process in the ER and Golgi apparatus, involving a large number of enzymes capable of fine-tuning structures. Changes in the localization of HS-modifying enzymes throughout the Golgi, rather than protein expression levels, were found to correlate with changes in the structure of HS. Following brefeldin A treatment, the HS-modifying enzymes localized preferentially in COPII vesicles and at the trans-Golgi. Further, shortly after treatment with heparin, the HS-modifying enzyme moved from cis to trans-Golgi, which coincided with increased HS trisulfated disaccharide content. Finally, it was shown that COPI subunits and Sec24 gene expression changed. Collectively, these findings highlight that the ER-Golgi dynamics of HS-modifying enzymes via vesicular trafficking processes are critical prerequisite for the complete delineation of HS biosynthesis.

cell biology↗