Method For Measuring Sub-Micron Facula At Single-Photon Level With Silicon Photomultiplier

Sub-micron faculae (light spots) at the single-photon level have important applications in many fields. This report demonstrates a method for measuring facula size at the sub-micron single-photon level indirectly. The developed method utilizes Silicon Photomultipliers (SiPMs) as the single-photon response detectors, combined with a nano-positioning stage. The approach involves one- or two-dimensional space scanning and a deconvolution operation, which enable evaluations of the size and spatial distribution of focused facula in a single-photon-level pulsed laser. The results indicate that the average full width at half maximum of the faculae is about 0.66 µm, which is close to the nominal resolution of the objective lens of the microscope (0.42 µm). The proposed method has two key advantages: (1) it can measure sub-micron facula at the single-photon level, and (2) the sub-micron facula can easily be aligned with the detector because the array area of the avalanche photodiode cells in SiPM is usually larger than one square millimeter, and there is no need to put an optical slit, knife edge, or pinhole in front of the detector. The method described herein is applicable in weak light facula detection related fields.

Properties of multi-vesicular release from rod photoreceptors support transmission of single photon responses

Vision under starlight requires rod photoreceptors to transduce and transmit single photon responses to the visual system. This remarkable sensitivity depends on a small voltage change reliably reducing glutamate release such that post-synaptic rod bipolar cells can robustly detect the signal. To transmit this small signal, we have found that rod vesicle release deviates strongly from a Poisson process under conditions that mimic darkness. Specifically, at their resting membrane potential in darkness, rods exhibit coordinated and regularly timed multivesicular release events. Each release event consisted of ∼17 vesicles and occurred 2-3 times more regularly than expected from a Poisson process. Hyperpolarizing rods to mimic the voltage change produced by a single photon response abruptly reduced the probability of multivesicular release nearly to zero with a rebound increase in release probability at stimulus offset. Simulations of these release dynamics indicate that this regularly timed, multivesicular release promotes transmission of single photon responses to post-synaptic neurons. Furthermore, the mechanism is efficient, requiring fewer vesicles to be released per second than uniquantal release governed by Poisson statistics.

Analytical Study of Front-End Circuits Coupled to Silicon Photomultipliers for Timing Performance Estimation under the Influence of Parasitic Components

Full exploitation of the intrinsic fast timing capabilities of analog silicon photomultipliers (SiPMs) requires suitable front-end electronics. Even a parasitic inductance of a few nH, associated to the interconnections between the SiPM and the preamplifier, can significantly degrade the steepness of the detector response, thus compromising the timing accuracy. In this work, we propose a simple analytic expression for the single-photon response of a SiPM coupled to the front-end electronics, as a function of the main parameters of the detector and the preamplifier, taking into account the parasitic inductance. The model is useful to evaluate the influence of each parameter of the system on the slope of its response and to guide the designer in the definition of the architecture and the specifications for the front-end electronics. The results provided by the model have been successfully compared with experimental measurements from a front-end circuit with variable configuration based on a bipolar junction transistor (BJT), coupled to a 3 × 3 mm2 SiPM stimulated by a fast-pulsed laser source.

Elementary response triggered by transducin in retinal rods

G protein-coupled receptor (GPCR) signaling is crucial for many physiological processes. A signature of such pathways is high amplification, a concept originating from retinal rod phototransduction, whereby one photoactivated rhodopsin molecule (Rho*) was long reported to activate several hundred transducins (GT*s), each then activating a cGMP-phosphodiesterase catalytic subunit (GT*·PDE*). This high gain at the Rho*-to-GT* step has been challenged more recently, but estimates remain dispersed and rely on some nonintact rod measurements. With two independent approaches, one with an extremely inefficient mutant rhodopsin and the other with WT bleached rhodopsin, which has exceedingly weak constitutive activity in darkness, we obtained an estimate for the electrical effect from a single GT*·PDE* molecular complex in intact mouse rods. Comparing the single-GT*·PDE* effect to the WT single-photon response, both in Gcaps−/− background, gives an effective gain of only ∼12–14 GT*·PDE*s produced per Rho*. Our findings have finally dispelled the entrenched concept of very high gain at the receptor-to-G protein/effector step in GPCR systems.