Standardizing a method for functional assessment of neural networks in brain organoids.

During the last decade brain organoids have emerged as an attractive model system, allowing stem cells to be differentiated into complex 3D models, recapitulating many aspects of human brain development. Whilst many studies have analysed anatomical and cytoarchitectural characteristics of organoids, their functional characterisation has been limited, and highly variable between studies. Standardised, consistent methods for recording functional activity are critical to providing a functional understanding of neuronal networks at the synaptic and network level that can yield useful information about functional network phenotypes in disease and healthy states. In this study we outline a detailed methodology for calcium imaging and Multi-Electrode Array (MEA) recordings in brain organoids. To illustrate the utility of these functional interrogation techniques in uncovering induced differences in neural network activity we applied various stimulating media protocols. We demonstrate overlapping information from the two modalities, with comparable numbers of active cells in the four treatment groups and an increase in synchronous behaviour in BrainPhys treated groups. Further development of analysis pipelines to reveal network level changes in brain organoids will enrich our understanding of network formation and perturbation in these structures, and aid in the future development of drugs that target neurological disorders at the network level.

Neurons Specifically Activated by Fear Learning in Lateral Amygdala Display Increased Synaptic Strength.

The lateral amygdala (LA) plays a critical role in the formation of fear-conditioned associative memories. Previous studies have used c-fos regulated expression to identify a spatially restricted population of neurons within the LA that is specifically activated by fear learning. These neurons are likely to be a part of a memory engram, but, to date, functional evidence for this has been lacking. We show that neurons within a spatially restricted region of the LA had an increase in both the frequency and amplitude of spontaneous postsynaptic currents (sPSC) when compared to neurons recorded from home cage control mice. We then more specifically addressed if this increased synaptic activity was limited to learning-activated neurons. Using a fos-tau-LacZ (FTL) transgenic mouse line, we developed a fluorescence-based method of identifying and recording from neurons activated by fear learning (FTL+ ) in acute brain slices. An increase in frequency and amplitude of sPSCs was observed in FTL+ neurons when compared to nonactivated FTL- neurons in fear-conditioned mice. No learning-induced changes were observed in the action potential (AP) input-output relationships. These findings support the idea that a discrete LA neuron population forms part of a memory engram through changes in synaptic connectivity.

Single-photon emitting diode in silicon carbide.

Electrically driven single-photon emitting devices have immediate applications in quantum cryptography, quantum computation and single-photon metrology. Mature device fabrication protocols and the recent observations of single defect systems with quantum functionalities make silicon carbide an ideal material to build such devices. Here, we demonstrate the fabrication of bright single-photon emitting diodes. The electrically driven emitters display fully polarized output, superior photon statistics (with a count rate of >300 kHz) and stability in both continuous and pulsed modes, all at room temperature. The atomic origin of the single-photon source is proposed. These results provide a foundation for the large scale integration of single-photon sources into a broad range of applications, such as quantum cryptography or linear optics quantum computing.

Ultra-high-density 3D DNA arrays within nanoporous biocompatible membranes for single-molecule-level detection and purification of circulating nucleic acids.

Extracellular nucleic acids freely circulating in blood and other physiologic fluids are important biomarkers for non-invasive diagnostics and early detection of cancer and other diseases, yet difficult to detect because they exist in very low concentrations and large volumes. Here we demonstrate a new broad-range sensor platform for ultrasensitive and selective detection of circulating DNA down to the single-molecule level. The biosensor is based on a chemically functionalized nanoporous diamond-like carbon (DLC) coated alumina membrane. The few nanometer-thick, yet perfect and continuous DLC-coating confers the chemical stability and biocompatibility of the sensor, allowing its direct application in biological conditions. The selective detection is based on complementary hybridization of a fluorescently-tagged circulating cancer oncomarker (a 21-mer nucleic acid) with covalently immobilized DNA on the surface of the membrane. The captured DNAs are detected in the nanoporous structure of the sensor using confocal scanning laser microscopy. The flow-through membrane sensor demonstrates broad-range sensitivity, spanning from 10(15) molecules per cm(2) down to single molecules, which is several orders of magnitude improvement compared to the flat DNA microarrays. Our study suggests that these flow-through type nanoporous sensors represent a new powerful platform for large volume sampling and ultrasensitive detection of different chemical biomarkers.

Temporal ringdown of silicon-on-insulator racetrack resonators.

Ring-down temporal measurements of silicon-on-insulator wire racetrack resonators are performed with 150 fs input pulses using a parametric process in a nonlinear crystal to gate and amplify the weak output pulses. We measure the cavity round trip time and the quality factor of these all-pass filters and find excellent agreement with continuous wave spectroscopic measurements as well as with an analytic model built using numerical solutions for the fully vectorial waveguide modes.

Uniformity of the lasing wavelength of heterogeneously integrated InP microdisk lasers on SOI.

We report a high lasing wavelength uniformity of optically pumped InP-based microdisk lasers processed with electron-beam lithography, heterogeneously integrated with adhesive bonding on silicon-on-insulator (SOI) waveguide circuits and evanescently coupled to an underlying waveguide. We study the continuous wave laser emission coupling out of the SOI via a grating coupler etched at one side of the waveguide, and demonstrate a standard deviation in lasing wavelength of nominally identical devices on the same chip lower than 500 pm. The deviation in the diameter of the microdisks as low as a few nanometers makes all-optical signal processing applications requiring cascadability possible.

Hybrid III-V semiconductor/silicon nanolaser.

Heterogeneous integration of III-V compound semiconductors on Silicon on Insulator is one the key technology for next-generation on-chip optical interconnects. In this context, the use of photonic crystals lasers represents a disruptive solution in terms of footprint, activation energy and ultrafast response. In this work, we propose and fabricate very compact laser sources integrated with a passive silicon waveguide circuitry. Using a subjacent Silicon-On-Insulator waveguide, the emitted light from a photonic crystal based cavity laser is efficiently captured. We study experimentally the evanescent wave coupling responsible for the funneling of the emitted light into the silicon waveguide mode as a function of the hybrid structure parameters, showing that 90% of coupling efficiency is possible.

Thermo-optical dynamics in an optically pumped Photonic Crystal nano-cavity.

Linear and non-linear thermo-optical dynamical regimes were investigated in a photonic crystal cavity. First, we have measured the thermal relaxation time in an InP-based nano-cavity with quantum dots in the presence of optical pumping. The experimental method presented here allows one to obtain the dynamics of temperature in a nanocavity based on reflectivity measurements of a cw probe beam coupled through an adiabatically tapered fiber. Characteristic times of 1.0+/-0.2 micros and 0.9+/-0.2 micros for the heating and the cooling processes were obtained. Finally, thermal dynamics were also investigated in a thermo-optical bistable regime. Switch-on/off times of 2 micros and 4 micros respectively were measured, which could be explained in terms of a simple non-linear dynamical representation.

Continuous-wave operation of photonic band-edge laser near 1.55 microm on silicon wafer.

We report on the continuous-wave operation of a band edge laser at room temperature near 1.55 mum in an InGaAs/InP photonic crystal. A flat dispersion band-edge photonic mode is used for surface normal operation. The photonic crystal slab is integrated onto a Silicon chip by means of Au/In bonding technology, which combines two advantages, efficient heat sinking and broad band reflectivity.

Direct observation of Bloch harmonics and negative phase velocity in photonic crystal waveguides.

The eigenfield distribution and the band structure of a photonic crystal waveguide have been measured with a phase-sensitive near-field scanning optical microscope. Bloch modes, which consist of more than one spatial frequency, are visualized in the waveguide. In the band structure, multiple Brillouin zones due to zone folding are observed, in which positive and negative dispersion is seen. The negative slopes are shown to correspond to a negative phase velocity but a positive group velocity. The lateral mode profile for modes separated by one reciprocal lattice vector is found to be different.

Real-space observation of ultraslow light in photonic crystal waveguides.

We show the real-space observation of fast and slow pulses propagating inside a photonic crystal waveguide by time-resolved near-field scanning optical microscopy. Local phase and group velocities of modes are measured. For a specific optical frequency we observe a localized pattern associated with a flat band in the dispersion diagram. During at least 3 ps, movement of this field is hardly discernible: its group velocity would be at most c/1000. The huge trapping times without the use of a cavity reveal new perspectives for dispersion and time control within photonic crystals.