William G. A. Brown

Gold‐Nanorod‐Assisted Near‐Infrared Stimulation of Primary Auditory Neurons

Infrared stimulation offers an alternative to electrical stimulation of neuronal tissue, with potential for direct, non-contact activation at high spatial resolution. Conventional methods of infrared neural stimulation (INS) rely on transient heating due to the absorption of relatively intense laser beams by water in the tissue. However, the water absorption also limits the depth of penetration of light in tissue. Therefore, the use of a near-infrared laser at 780 nm to stimulate cultured rat primary auditory neurons that are incubated with silica-coated gold nanorods (Au NRs) as an extrinsic absorber is investigated. The laser-induced electrical behavior of the neurons is observed using whole-cell patch clamp electrophysiology. The nanorod-treated auditory neurons (NR-ANs) show a significant increase in electrical activity compared with neurons that are incubated with non-absorbing silica-coated gold nanospheres and control neurons with no gold nanoparticles. The laser-induced heating by the nanorods is confirmed by measuring the transient temperature increase near the surface of the NR-ANs with an open pipette electrode. These findings demonstrate the potential to improve the efficiency and increase the penetration depth of INS by labeling nerves with Au NRs and then exposing them to infrared wavelengths in the water window of tissue.

Modeling of light absorption in tissue during infrared neural stimulation

A Monte Carlo model has been developed to simulate light transport and absorption in neural tissue during infrared neural stimulation (INS). A range of fiber core sizes and numerical apertures are compared illustrating the advantages of using simulations when designing a light delivery system. A range of wavelengths, commonly used for INS, are also compared for stimulation of nerves in the cochlea, in terms of both the energy absorbed and the change in temperature due to a laser pulse. Modeling suggests that a fiber with core diameter of 200 μm and NA=0.22 is optimal for optical stimulation in the geometry used and that temperature rises in the spiral ganglion neurons are as low as 0.1°C. The results show a need for more careful experimentation to allow different proposed mechanisms of INS to be distinguished.

Modeling of the temporal effects of heating during infrared neural stimulation.

Abstract. A model of infrared neural stimulation (INS) has been developed to allow the temporal characteristics of different stimulation parameters and geometries to be better understood. The model uses a finite element approach to solve the heat equation and allow detailed analysis of heat during INS with both microsecond and millisecond laser pulses. When compared with experimental data, the model provides insight into the mechanisms behind INS. In particular, the analysis suggests that there may be two broad regimes of INS: the process tends to be limited by the total pulse energy for pulse lengths below 100 μs, while the temperature gradient with respect to time becomes more important above 100 μs.

Nanoparticle-enhanced infrared neural stimulation.

OBJECTIVE Recent research has demonstrated that nerves can be stimulated by transient heating associated with the absorption of infrared light by water in the tissue. There is a great deal of interest in using this technique in neural prostheses, due to the potential for increased localization of the stimulus and minimization of contact with the tissue. However, thermal modelling suggests that the full benefits of increased localization may be reduced by cumulative heating effects when multiple stimulus sites and/or high repetition rates are used. APPROACH Here we review recent in vitro and in vivo results suggesting that the transient heating associated with plasmon absorption in gold nanorods can also be used to stimulate nerves. MAIN RESULTS Patch clamp experiments on cultured spiral ganglion neurons exhibited action potentials when exposed to 780 nm light at the plasmon absorption peak, while the amplitude of compound action potentials in the rat sciatic nerve were increased by laser irradiation of gold nanorods in the vicinity of the plasma membrane. Similarly, calcium imaging studies of NG108-15 neuronal cells incubated with Au nanorods revealed an increased level of intracellular calcium activity synchronized with laser exposure. SIGNIFICANCE Given that the plasmon absorption peak of gold nanorods can be matched with the transparency window of biological tissues, these results demonstrate that nanorod absorbers hold great promise to enhance the process of infrared neural stimulation for future applications in neural prostheses and fundamental studies in neuroscience.

Effect of phase mask alignment on fiber Bragg grating spectra at harmonics of the Bragg wavelength

Effects of fabrication conditions on the double-peak structure observed in fiber Bragg gratings at harmonics of the Bragg wavelength were investigated, showing that slight variations in the alignment of the phase mask can affect the grating spectra significantly. A single peak occurs only when the incident beam direction is perfectly normal with respect to the fiber.

Anomalous dispersion and negative group velocity in a coherence-free cold atomic medium

We have observed the propagation of an approximately 35 ns long light pulse with a negative group velocity through a laser-cooled 85Rb atomic medium. The anomalous dispersion results from linear atom-light interaction and is unrelated to long-lived ground-state coherences often associated with fast light in atomic media. The observed negative group velocity (−c/360) in the Rb magneto-optical trap for a pulse attenuated by less than 50% is in good agreement with the value of dispersion measured independently by a rf heterodyne method. The spectral region of anomalous dispersion is between 15 and 40 MHz, which is an order of magnitude wider than that typically associated with ground-state coherences.

Whole Cell Patch Clamp for Investigating the Mechanisms of Infrared Neural Stimulation

It has been demonstrated in recent years that pulsed, infrared laser light can be used to elicit electrical responses in neural tissue, independent of any further modification of the target tissue. Infrared neural stimulation has been reported in a variety of peripheral and sensory neural tissue in vivo, with particular interest shown in stimulation of neurons in the auditory nerve. However, while INS has been shown to work in these settings, the mechanism (or mechanisms) by which infrared light causes neural excitation is currently not well understood. The protocol presented here describes a whole cell patch clamp method designed to facilitate the investigation of infrared neural stimulation in cultured primary auditory neurons. By thoroughly characterizing the response of these cells to infrared laser illumination in vitro under controlled conditions, it may be possible to gain an improved understanding of the fundamental physical and biochemical processes underlying infrared neural stimulation.

Comparison of spectra and images of Bragg gratings written in three different optical fibres

Optical fibre Bragg gratings were fabricated in 3 different fibres, and their spectra at 3 harmonics were measured and cross-compared with images of their internal complex refractive index profiles obtained via differential interface contrast imaging.

Time-dependent activity of primary auditory neurons in the presence of neurotrophins and antibiotics

Abstract In vitro cultures provide a valuable tool in studies examining the survival, morphology and function of cells in the auditory system. Primary cultures of primary auditory neurons have most notably provided critical insights into the role of neurotrophins in cell survival and morphology. Functional studies have also utilized in vitro models to study neuronal physiology and the ion channels that dictate these patterns of activity. Here we examine what influence time‐in‐culture has on the activity of primary auditory neurons, and how this affects our interpretation of neurotrophin and antibiotic‐mediated effects in this population. Using dissociated cell culture we analyzed whole‐cell patch‐clamp recordings of spiral ganglion neurons grown in the presence or absence of neurotrophins and/or penicillin and streptomycin for 1–3 days in vitro. Firing threshold decreased, and both action potential number and latency increased over time regardless of treatment, whilst input resistance was lowest where neurotrophins were present. Differences in firing properties were seen with neurotrophin concentration but were not consistently maintained over the 3 days in vitro. The exclusion of antibiotics from culture media influenced most firing properties at 1 day in vitro in both untreated and neurotrophin‐treated conditions. The only difference still present at 3 days was an increase in input resistance in neurotrophin‐treated neurons. These results highlight the potential of neurotrophins and antibiotics to influence neural firing patterns in vitro in a time‐dependent manner, and advise the careful consideration of their impact on SGN function in future studies. HighlightsThe electrophysiological properties of SGNs are not maintained with time in vitro.Antibiotic modulation of SGN activity and passive properties is time‐dependent.Our results indicate shorter time in vitro better reflects native SGN activity.

The role of membrane capacitance and ion channels in the response of primary auditory neurons to infrared light (Conference Presentation)

Infrared light can be used to modulate the activity of neuronal cells with broad generality and without any need for exogenous materials. The action potential response has been shown to be associated with heating due to the absorption of light by water in and around the illuminated tissues, which gives rise to at least two distinct processes: namely, the temperature pulses cause depolarizing capacitive currents due to an intramembrane thermo-mechanical effect, and in addition, temperature-sensitive TRPV ion channels (and likely, voltage-gated channels) drive additional membrane depolarization. However, substantial differences between the activation threshold of primary auditory neurons ( 300 mJ/cm^2) in vivo have generated some controversy in the field. A temperature-dependent Hodgkin-Huxley type model, which combines capacitive currents and the experimentally-derived characteristics of voltage-gated potassium and sodium ion channels in primary auditory neurons, was used to accurately explain the in vitro response to 1870 nm infrared illumination. TRPV channels do not make a significant contribution in this case, suggesting that the detailed mechanism of the neuronal response to infrared light is dependent on the specific cell type. Furthermore, based on this detailed understanding of the cell behaviour, it is shown that action potentials cannot be generated at safe laser power levels. This suggests that the previously reported response of the auditory system to infrared stimulation in vivo might arise from a different mechanism, and calls into question the potential usefulness of the effect for auditory prostheses.