Abstract

In infrared neural stimulation (INS), laser-evoked thermal transients are used to generate small depolarising currents in neurons. The laser exposure poses a moderate risk of thermal damage to the target neuron. Indeed, exogenous methods of neural stimulation often place the target neurons under stressful non-physiological conditions, which can hinder ordinary neuronal function and hasten cell death. Therefore, quantifying the exposure-dependent probability of neuronal damage is essential for identifying safe operating limits of INS and other interventions for therapeutic and prosthetic use. Using patch-clamp recordings in isolated spiral ganglion neurons, we describe a method for determining the dose-dependent damage probabilities of individual neurons in response to both acute and cumulative infrared exposure parameters based on changes in injection current. The results identify a local thermal damage threshold at approximately 60 °C, which is in keeping with previous literature and supports the claim that damage during INS is a purely thermal phenomenon. In principle this method can be applied to any potentially injurious stimuli, allowing for the calculation of a wide range of dose-dependent neural damage probabilities. Unlike histological analyses, the technique is well-suited to quantifying gradual neuronal damage, and critical threshold behaviour is not required.

Highlights

  • Artificial stimulation of neurons constitutes the backbone of investigative electrophysiology and translational neuroscience

  • Cell damage probability probability of cell damage (Pd) exhibited no dependence on peak laser power in both the room temperature (22.4 °C) and elevated temperature (30.4 °C) conditions, up to a maximum of 1.5 W (Fig. 3(a, b))

  • Our results suggest that spiral ganglion neurons (SGNs) can safely tolerate temperature gradients of at least 14 °C ms−1 during infrared exposure, and appears to rule out the possibility of cellular damage due to phototoxic interactions, which would be expected to scale with photon flux

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Summary

Introduction

Artificial stimulation of neurons constitutes the backbone of investigative electrophysiology and translational neuroscience. Evaluating the safe operating parameters of these techniques is essential for their clinical translation, and typically relies on histological analysis of the target cells or fluorescence staining using dye assays ex situ. These detection methods are usually destructive, making them unsuitable for observing the time course of neural degradation. Patch-clamp techniques for intracellular electrophysiological recording can provide real-time measurements of a neuron’s membrane potential, impedance and transmembrane current. Membrane impedance can provide a useful estimate of stimulus-dependent damage probability over the course of an experiment

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