Anion and cation permeability of the mouse tmem16f calcium-activated channel

TMEM16F is involved in several physiological processes, such as blood coagulation, bone development and virus infections. This protein acts both as a Ca2+-dependent phospholipid scram-blase and a Ca2+-activated ion channel but several studies have reported conflicting results about the ion selectivity of the TMEM16F-mediated current. Here, we have performed a detailed side-by-side comparison of the ion selectivity of TMEM16F using the whole-cell and inside-out excised patch configurations to directly compare the results. In inside-out configuration, Ca2+-dependent activation was fast and the TMEM16F-mediated current was activated in a few milliseconds, while in whole-cell recordings full activation required several minutes. We determined the relative permeability between Na+ and Cl¯ (PNa /PCl ) using the dilution method in both configurations. The TMEM16F-mediated current was highly nonselective, but there were differences depending on the configuration of the recordings. In whole-cell recordings, PNa /PCl was approximately 0.5, indicating a slight preference for Cl¯ permeation. In contrast, in inside-out experiments the TMEM16F channel showed a higher permeability for Na+ with PNa /PCl reaching 3.7. Our results demonstrate that the time dependence of Ca2+ activation and the ion selectivity of TMEM16F depend on the recording configuration

Note: Comparison between a prism-based and an acousto-optic tunable filter-based spectrometer for diffusive media

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Studying the evolution in time of bimetallic nanoparticles morphology by Cyclic Voltammetry

Over the last decades, bimetallic nanosized systems have attracted more and more interest thanks to their novel optical, catalytic, magnetic, and sensing properties, often different from the ones of their monometallic counterparts. Studies directed towards the size, shape, composition, and functionalization of the bimetallic nanoparticles are optimized to design sophisticated materials for the intended applications. Considering these facts, it is important to evaluate not only the type and the quantity of the two metals involved, but also their morphological distribution (e.g. alloy or core-shell). Characterization techniques normally used to investigate bimetallic systems are HR-TEM and EXAFS, very expensive and not so easily accessible. Recently, electrochemistry has been employed as alternative or complementary, low-cost, efficient technique with very promising results, allowing the discrimination between alloyed and perfect or defective core-shell systems after their synthesis. A further achievement is the possibility to follow step by step the formation morphology of these nanomaterials during their synthetic procedure. In the present work, we present a study on Au-Pt bimetallic nanoparticles, in form of alloy or core-shell. Cyclovoltammetry (CV) is used as a fast, low-cost and simple screening technique to distinguish the general composition of the sample and to understand the evolution in time of the systems morphology during their synthesis. An additional advantage is the possibility to conduct the study of the material simply in liquid form, without the need of using solid supports, as normally required by other characterization techniques. Interesting results are obtained for Au-based bimetallic samples, gaining information in accordance with TEM images and EXAFS spectra. This fact moves the interest towards the study of other bimetallic systems, to be used in catalytic, electrocatalytic and electroanalytical applications

Time resolved diffuse optical spectroscopy with geometrically accurate models for bulk parameter recovery

A novel straightforward, accessible and efficient approach is presented for performing hyperspectral time-domain diffuse optical spectroscopy to determine the optical properties of samples accurately using geometry specific models. To allow bulk parameter recovery from measured spectra, a set of libraries based on a numerical model of the domain being investigated is developed as opposed to the conventional approach of using an analytical semi-infinite slab approximation, which is known and shown to introduce boundary effects. Results demonstrate that the method improves the accuracy of derived spectrally varying optical properties over the use of the semi-infinite approximation

Spectrally Resolved Single-Photon Timing of Silicon Photomultipliers for Time-Domain Diffuse Spectroscopy

We characterized the single-photon timing response function of various silicon photomultipliers (SiPMs) over a broad (500-1100 nm) spectral range. We selected two SiPM manufacturers, and we investigated two active areas, i.e., a small (1-1.69 mm2 and a large (9 mm2) one, for each of them. We demonstrate that selected SiPMs are suitable for time-resolved diffuse optics (DO) applications where a very large detection area and sensitivity down to single photons are crucial to detecting the very faint return signal from biological tissues, like the brain, thus allowing replacement of photomultiplier tubes and opening the way to a novel generation of DO multichannel instrumentation. Due to our custom front-end electronics, we show the world's best single-photon timing resolution for SiPMs, namely, 57-ps full-width at half maximum for Hamamatsu 1.69 mm2 and 115 ps for Excelitas 9 mm2. Even further, we provide a thorough spectral investigation of the full single-photon timing response function, also detailing diffusion tails' time constants and dynamic range. The achieved insight and the reported performance open the way to a widespread diffusion of SiPMs not just in many-photon regimes (e.g., PET) but at single-photon counting regimes like DO as well

Time-resolved single-photon detection module based on silicon photomultiplier: A novel building block for time-correlated measurement systems

We present the design and preliminary characterization of the first detection module based on Silicon Photomultiplier (SiPM) tailored for single-photon timing applications. The aim of this work is to demonstrate, thanks to the design of a suitable module, the possibility to easily exploit SiPM in many applications as an interesting detector featuring large active area, similarly to photomultipliers tubes, but keeping the advantages of solid state detectors (high quantum efficiency, low cost, compactness, robustness, low bias voltage, and insensitiveness to magnetic field). The module integrates a cooled SiPM with a total photosensitive area of 1 mm2 together with the suitable avalanche signal read-out circuit, the signal conditioning, the biasing electronics, and a Peltier cooler driver for thermal stabilization. It is able to extract the single-photon timing information with resolution better than 100 ps full-width at half maximum. We verified the effective stabilization in response to external thermal perturbations, thus proving the complete insensitivity of the module to environment temperature variations, which represents a fundamental parameter to profitably use the instrument for real-field applications. We also characterized the single-photon timing resolution, the background noise due to both primary dark count generation and afterpulsing, the single-photon detection efficiency, and the instrument response function shape. The proposed module can become a reliable and cost-effective building block for time-correlated single-photon counting instruments in applications requiring high collection capability of isotropic light and detection efficiency (e.g., fluorescence decay measurements or time-domain diffuse optics systems)

Expression pattern of Stomatin-domain proteins in the peripheral olfactory system

Recent data show that Stomatin-like protein 3 (STOML3), a member of the stomatin-domain family, is expressed in the olfactory sensory neurons (OSNs) where it modulates both spontaneous and evoked action potential firing. The protein family is constituted by other 4 members (besides STOML3): STOM, STOML1, STOML2 and podocin. Interestingly, STOML3 with STOM and STOML1 are expressed in other peripheral sensory neurons: dorsal root ganglia. In here, they functionally interact and modulate the activity of the mechanosensitive Piezo channels and members of the ASIC family. Therefore, we investigated whether STOM and STOML1 are expressed together with STOML3 in the OSNs and whether they could interact. We found that all three are indeed expressed in ONSs, although STOML1 at very low level. STOM and STOML3 share a similar expression pattern and STOML3 is necessary for STOM to properly localize to OSN cilia. In addition, we extended our investigation to podocin and STOML2, and while the former is not expressed in the olfactory system, the latter showed a peculiar expression pattern in multiple cell types. In summary, we provided a first complete description of stomatin-domain protein family in the olfactory system, highlighting the precise compartmentalization, possible interactions and, finally, their functional implications

Bandpass Effects in Time-Resolved Diffuse Spectroscopy

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Novel approaches to photon detection and timing for 7-wavelength time domain optical mammography

An 8-channel Silicon Photomultiplier probe and a Time-to-Digital Converter are used to build a higher-throughput, cheaper and compact detection chain for time-resolved optical mammography as compared with conventional PhotoMultiplier Tubes and Time-Correlated Single-Photon Counting boards, still providing comparable performance in the estimation of optical properties, but with higher optical responsivity

Interactions between permeation and gating in the TMEM16B/anoctamin2 calcium-activated chloride channel

At least two members of the TMEM16/anoctamin family, TMEM16A (also known as anoctamin1) and TMEM16B (also known as anoctamin2), encode Ca2+-activated Cl- channels (CaCCs), which are found in various cell types and mediate numerous physiological functions. Here, we used whole-cell and excised inside-out patch-clamp to investigate the relationship between anion permeation and gating, two processes typically viewed as independent, in TMEM16B expressed in HEK 293T cells. The permeability ratio sequence determined by substituting Cl- with other anions (PX/PCl) was SCN- > I- > NO3- > Br- > Cl- > F- > gluconate. When external Cl- was substituted with other anions, TMEM16B activation and deactivation kinetics at 0.5 μM Ca2+ were modified according to the sequence of permeability ratios, with anions more permeant than Cl- slowing both activation and deactivation and anions less permeant than Cl- accelerating them. Moreover, replacement of external Cl- with gluconate, or sucrose, shifted the voltage dependence of steady-state activation (G-V relation) to more positive potentials, whereas substitution of extracellular or intracellular Cl- with SCN- shifted G-V to more negative potentials. Dose-response relationships for Ca2+ in the presence of different extracellular anions indicated that the apparent affinity for Ca2+ at +100 mV increased with increasing permeability ratio. The apparent affinity for Ca2+ in the presence of intracellular SCN- also increased compared with that in Cl-. Our results provide the first evidence that TMEM16B gating is modulated by permeant anions and provide the basis for future studies aimed at identifying the molecular determinants of TMEM16B ion selectivity and gating. © 2014 Betto et al