Layer-specific stimulations of parvalbumin-positive cortical interneurons in mice entrain brain rhythms to different frequencies.

Neocortical interneurons provide inhibition responsible for organizing neuronal activity into brain oscillations that subserve cognitive functions such as memory, attention, or prediction. However, the interneuronal contribution to the entrainment of neocortical oscillations within and across different cortical layers was not described. Here, using layer-specific optogenetic stimulations with micro-Light-Emitting Diode arrays, directed toward parvalbumin-expressing (PV) interneurons in non-anesthetized awake mice, we found that supragranular layer stimulations of PV neurons were most efficient at entraining supragranular local field potential (LFP) oscillations at gamma frequencies (γ: 25-80 Hz), whereas infragranular layer stimulation of PV neurons better entrained the LFP at delta (δ: 2-5 Hz) and theta (θ: 6-10 Hz) frequencies. At the level of neuronal action potential activity, we observed that supragranular neurons better followed the imposed PV stimulation rhythm than their infragranular counterparts at most frequencies when the stimulation was delivered in their respective layer. Moreover, the neuronal entrainment evoked by local stimulation could propagate across layers, though with a lesser impact when the stimulation occurs in deep layers, suggesting a direction-specific laminar propagation. These results establish a layer-based framework for oscillations to entrain the primary somatosensory cortex in awake conditions.

Hybrid intracerebral probe with integrated bare LED chips for optogenetic studies.

This article reports on the development, i.e., the design, fabrication, and validation of an implantable optical neural probes designed for in vivo experiments relying on optogenetics. The probes comprise an array of ten bare light-emitting diode (LED) chips emitting at a wavelength of 460 nm and integrated along a flexible polyimide-based substrate stiffened using a micromachined ladder-like silicon structure. The resulting mechanical stiffness of the slender, 250-µm-wide, 65-µm-thick, and 5- and 8-mm-long probe shank facilitates its implantation into neural tissue. The LEDs are encapsulated by a fluropolymer coating protecting the implant against the physiological conditions in the brain. The electrical interface to the external control unit is provided by 10-µm-thick, highly flexible polyimide cables making the probes suitable for both acute and chronic in vivo experiments. Optical and electrical properties of the probes are reported, as well as their in vivo validation in acute optogenetic studies in transgenic mice. The depth-dependent optical stimulation of both excitatory and inhibitory neurons is demonstrated by altering the brain activity in the cortex and the thalamus. Local network responses elicited by 20-ms-long light pulses of different optical power (20 µW and 1 mW), as well as local modulation of single unit neuronal activity to 1-s-long light pulses with low optical intensity (17 µW) are presented. The ability to modulate neural activity makes these devices suitable for a broad variety of optogenetic experiments.

Compact Optical Neural Probes With Up to 20 Integrated Thin-Film µLEDs Applied in Acute Optogenetic Studies.

This paper reports on the development, characterization and in vivo validation of compact optical neural probes. These novel intracerebral devices comprise micro light-emitting diodes ( µLEDs) integrated along their slender probe shanks with up to 20 µLEDs per device. Blue light with a peak wavelength of 455 nm is emitted from circular apertures 100 µm in diameter. The µLEDs are structured on GaN-on-sapphire wafers and subsequently transferred onto silicon (Si) carrier wafers. The wafer-scale transfer process provides the opportunity to process the functional GaN layer stack from both sides and hence enables maximizing the efficiency of the µLEDs. Combined with standard MEMS fabrication processes for Si, linear µLED arrays with small inter- µLED distances are achieved on thin probe shanks with cross-sections measuring [Formula: see text]. Devices are interconnected using highly flexible polyimide cables in order to mechanically decouple them from the peripheral electronics during in vivo experiments. Assembled probes emit a peak optical radiant flux of 440 µW (emittance 56 mW mm -2) at 5 mA driving current. Thermal characterization of test probes reveals a temperature increase of 1.5 K measured using an integrated thermistor. Electrical functionality stress tests have been carried out to evaluate the device passivation against the physiological environment. It is estimated to endure at least 48 h during continuously pulsed µLED operation. A compact driving circuitry enables low-noise µLED operation in in vivo optogenetic experiments. The radiant flux necessary to elicit an acceptable neuronal response is determined between 1.36 µW and 17.5 µW. Probe validation successfully demonstrates the layer-specific stimulation in the cortex in multiple in vivo trials.

Multichannel optogenetic stimulation of the auditory pathway using microfabricated LED cochlear implants in rodents.

When hearing fails, electrical cochlear implants (eCIs) provide the brain with auditory information. One important bottleneck of CIs is the poor spectral selectivity that results from the wide current spread from each of the electrode contacts. Optical CIs (oCIs) promise to make better use of the tonotopic order of spiral ganglion neurons (SGNs) inside the cochlea by spatially confined stimulation. Here, we established multichannel oCIs based on light-emitting diode (LED) arrays and used them for optical stimulation of channelrhodopsin (ChR)-expressing SGNs in rodents. Power-efficient blue LED chips were integrated onto microfabricated 15-µm-thin polyimide-based carriers comprising interconnecting lines to address individual LEDs by a stationary or mobile driver circuitry. We extensively characterized the optoelectronic, thermal, and mechanical properties of the oCIs and demonstrated stability over weeks in vitro. We then implanted the oCIs into ChR-expressing rats and gerbils, and characterized multichannel optogenetic SGN stimulation by electrophysiological and behavioral experiments. Improved spectral selectivity was directly demonstrated by recordings from the auditory midbrain. Long-term experiments in deafened ChR-expressing rats and in nontreated control animals demonstrated specificity of optogenetic stimulation. Behavioral studies on animals carrying a wireless oCI sound processor revealed auditory percepts. This study demonstrates hearing restoration with improved spectral selectivity by an LED-based multichannel oCI system.

Invasive Optical Pacing in Perfused, Optogenetically Modified Mouse Heart Using Stiff Multi-LED Optical Probes.

We present the first invasive use of a stiff, multiLED optical probe for intramural optical stimulation of cardiac tissue. We demonstrate that optical pacing is possible with high spatial and temporal resolution in transgenic mice expressing channelrhodopsin-2. The technical implementation of this study builds on optical probes recently developed and tested ex vivo in cerebral tissue of mice. The probes comprise LEDs integrated on flexible substrates stiffened by silicon-based MEMS structures enabling the successful penetration into the cardiac tissue. The probe technology is extended to allow dual-sided illumination for directional tissue stimulation. Implantation trials affirm the ability to optically pace the isolated perfused heart at stimulation frequencies between 4Hz and 12Hz with experimentally determined emittance levels of 10mW mm-2 Rapid activation of two distant LEDs could reliably be used to induce short runs of ventricular fibrillation, while simultaneous activation of all LEDs allowed termination of re-entrant rhythm disturbances (optical defibrillation). Thus, spatially-resolved intramural pacing and rhythm control of the isolated heart is possible using stiff, multi-LED optical probes.

Heterogeneous 3D optrode with variable spatial resolution for optogenetic stimulation and electrophysiological recording.

We report on the concept, development, and geometrical, optical as well as electrical characterization of the first three-dimensional (3D) optrode. This new device allows to optically interact with neuronal cells and simultaneously record their response with a high spatial resolution. Our design is based on a single-shank optical stimulation component and a multi-shank recording probe stacked together in a delicate assembly process. The electrical connection of both components is ensured by using flexible polyimide (PI) ribbon cables. The highly accurate relative positioning and precise alignment of the optical and electrical components in 3D with an optical output power at 460 nm well above 5 mW/mm2 and an all-electrical interface makes this device a promising tool for optogenetic experiments in neuroscientific research.