Efficient Synthesis of a Schiff Base Copper(II) Complex Using a Microfluidic Device.

The efficient synthesis of amino acid Schiff base copper(II) complexes using a microfluidic device was successfully achieved. Schiff bases and their complexes are remarkable compounds due to their high biological activity and catalytic function. Conventionally, products are synthesized under reaction conditions of 40 °C for 4 h using a beaker-based method. However, in this paper, we propose using a microfluidic channel to enable quasi-instantaneous synthesis at room temperature (23 °C). The products were characterized using UV-Vis, FT-IR, and MS spectroscopy. The efficient generation of compounds using microfluidic channels has the potential to significantly contribute to the efficiency of drug discovery and material development due to high reactivity.

Microfluidic cell engineering on high-density microelectrode arrays for assessing structure-function relationships in living neuronal networks.

Neuronal networks in dissociated culture combined with cell engineering technology offer a pivotal platform to constructively explore the relationship between structure and function in living neuronal networks. Here, we fabricated defined neuronal networks possessing a modular architecture on high-density microelectrode arrays (HD-MEAs), a state-of-the-art electrophysiological tool for recording neural activity with high spatial and temporal resolutions. We first established a surface coating protocol using a cell-permissive hydrogel to stably attach a polydimethylsiloxane microfluidic film on the HD-MEA. We then recorded the spontaneous neural activity of the engineered neuronal network, which revealed an important portrait of the engineered neuronal network-modular architecture enhances functional complexity by reducing the excessive neural correlation between spatially segregated modules. The results of this study highlight the impact of HD-MEA recordings combined with cell engineering technologies as a novel tool in neuroscience to constructively assess the structure-function relationships in neuronal networks.

Contribution of AMPA and NMDA receptors in the spontaneous firing patterns of single neurons in autaptic culture.

Single neurons in an autaptic culture exhibit various types of firing pattern with different firing durations and rhythms. However, a neuron with autapses has often been modeled as an oscillator providing a monotonic firing pattern with a constant periodicity because of the lack of a mathematical model. In the work described in this study, we use computational simulation and whole-cell patch-clamp recording to elucidate and model the mechanism by which such neurons generate various firing pattens. In the computational simulation, three types of spontaneous firing pattern, i.e., short, long-lasting, and periodic burst firing patterns are realized by changing the combination ratio of N-methyl-d-aspartate (NMDA) to α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) conductance. These three types of firing patterns are also observed in the experiments where neurons are cultured in isolation on micropatterned substrates. Using the AMPA and NMDA current models, we discuss that, in principle, autapses can regulate rhythmicity and information selection in neuronal networks.

[Creation of Stereo Color Anatomical Charts Showing Nerve Fibers in the Cerebral Lobe and Gyrus of the Brain for Use in the MR Examination].

In anatomical charts in conventional books, the pathways of nerve fibers are drawn in illustrations. Conversely, with diffusion tensor tractography (DTT), we can visually understand the trajectory of nerve fibers through color. We created a stereo color anatomical chart of the nerve fibers that can be used for magnetic resonance (MR) examination to diagnose the pathway of nerve fibers and that can be used to explain the results of MR examination to visually understand how nerve fiber information is transmitted from the frontal lobe, parietal lobe, occipital lobe, temporal lobe, cerebellar lobe, and cerebral cortex.

Triple nitrogen-vacancy centre fabrication by C5N4Hn ion implantation.

Quantum information processing requires quantum registers based on coherently interacting quantum bits. The dipolar couplings between nitrogen vacancy (NV) centres with nanometre separation makes them a potential platform for room-temperature quantum registers. The fabrication of quantum registers that consist of NV centre arrays has not advanced beyond NV pairs for several years. Further scaling up of coupled NV centres by using nitrogen implantation through nanoholes has been hampered because the shortening of the separation distance is limited by the nanohole size and ion straggling. Here, we demonstrate the implantation of C5N4Hn from an adenine ion source to achieve further scaling. Because the C5N4Hn ion may be regarded as an ideal point source, the separation distance is solely determined by straggling. We successfully demonstrate the fabrication of strongly coupled triple NV centres. Our method may be extended to fabricate small quantum registers that can perform quantum information processing at room temperature.

Room Temperature Resonant Photocurrent in an Erbium Low-Doped Silicon Transistor at Telecom Wavelength.

An erbium-doped silicon transistor prepared by ion implantation and co-doped with oxygen is investigated by photocurrent generation in the telecommunication range. The photocurrent is explored at room temperature as a function of the wavelength by using a supercontinuum laser source working in the µW range. The 1-µm² transistor is tuned to involve in the transport only those electrons lying in the Er-O states. The spectrally resolved photocurrent is characterized by the typical absorption line of erbium and the linear dependence of the signal over the impinging power demonstrates that the Er-doped transistor is operating far from saturation. The relatively small number of estimated photoexcited atoms (≈ 4 × 10 4 ) makes Er-dpoed silicon potentially suitable for designing resonance-based frequency selective single photon detectors at 1550 nm.

GeVn complexes for silicon-based room-temperature single-atom nanoelectronics.

We propose germanium-vacancy complexes (GeVn) as a viable ingredient to exploit single-atom quantum effects in silicon devices at room temperature. Our predictions, motivated by the high controllability of the location of the defect via accurate single-atom implantation techniques, are based on ab-initio Density Functional Theory calculations within a parameterfree screened-dependent hybrid functional scheme, suitable to provide reliable bandstructure energies and defect-state wavefunctions. The resulting defect-related excited states, at variance with those arising from conventional dopants such as phosphorous, turn out to be deep enough to ensure device operation up to room temperature and exhibit a far more localized wavefunction.

Impact of modular organization on dynamical richness in cortical networks.

As in many naturally formed networks, the brain exhibits an inherent modular architecture that is the basis of its rich operability, robustness, and integration-segregation capacity. However, the mechanisms that allow spatially segregated neuronal assemblies to swiftly change from localized to global activity remain unclear. Here, we integrate microfabrication technology with in vitro cortical networks to investigate the dynamical repertoire and functional traits of four interconnected neuronal modules. We show that the coupling among modules is central. The highest dynamical richness of the network emerges at a critical connectivity at the verge of physical disconnection. Stronger coupling leads to a persistently coherent activity among the modules, while weaker coupling precipitates the activity to be localized solely within the modules. An in silico modeling of the experiments reveals that the advent of coherence is mediated by a trade-off between connectivity and subquorum firing, a mechanism flexible enough to allow for the coexistence of both segregated and integrated activities. Our results unveil a new functional advantage of modular organization in complex networks of nonlinear units.

1.54 µm photoluminescence from Er:Ox centers at extremely low concentration in silicon at 300 K.

The demand for single photon emitters at λ=1.54 µm, which follows from the consistent development of quantum networks based on optical fiber technologies, makes Er:Ox centers in Si a viable resource, thanks to the I13/24→I415/2 optical transition of Er3+. While its implementation in high-power applications is hindered by the extremely low emission rate, the study of such systems in the low concentration regime remains relevant for quantum technologies. In this Letter, we explore the room-temperature photoluminescence at the telecomm wavelength from very low implantation doses of Er:Ox in Si. The lower-bound number of optically active Er atoms detected is of the order of 102, corresponding to a higher-bound value for the emission rate per individual ion of about 104 s-1.

Live-Cell, Label-Free Identification of GABAergic and Non-GABAergic Neurons in Primary Cortical Cultures Using Micropatterned Surface.

Excitatory and inhibitory neurons have distinct roles in cortical dynamics. Here we present a novel method for identifying inhibitory GABAergic neurons from non-GABAergic neurons, which are mostly excitatory glutamatergic neurons, in primary cortical cultures. This was achieved using an asymmetrically designed micropattern that directs an axonal process to the longest pathway. In the current work, we first modified the micropattern geometry to improve cell viability and then studied the axon length from 2 to 7 days in vitro (DIV). The cell types of neurons were evaluated retrospectively based on immunoreactivity against GAD67, a marker for inhibitory GABAergic neurons. We found that axons of non-GABAergic neurons grow significantly longer than those of GABAergic neurons in the early stages of development. The optimal threshold for identifying GABAergic and non-GABAergic neurons was evaluated to be 110 µm at 6 DIV. The method does not require any fluorescence labelling and can be carried out on live cells. The accuracy of identification was 98.2%. We confirmed that the high accuracy was due to the use of a micropattern, which standardized the development of cultured neurons. The method promises to be beneficial both for engineering neuronal networks in vitro and for basic cellular neuroscience research.

Size-dependent regulation of synchronized activity in living neuronal networks.

We study the effect of network size on synchronized activity in living neuronal networks. Dissociated cortical neurons form synaptic connections in culture and generate synchronized spontaneous activity within 10 days in vitro. Using micropatterned surfaces to extrinsically control the size of neuronal networks, we show that synchronized activity can emerge in a network as small as 12 cells. Furthermore, a detailed comparison of small (∼20 cells), medium (∼100 cells), and large (∼400 cells) networks reveal that synchronized activity becomes destabilized in the small networks. A computational modeling of neural activity is then employed to explore the underlying mechanism responsible for the size effect. We find that the generation and maintenance of the synchronized activity can be minimally described by: (1) the stochastic firing of each neuron in the network, (2) enhancement in the network activity in a positive feedback loop of excitatory synapses, and (3) Ca-dependent suppression of bursting activity. The model further shows that the decrease in total synaptic input to a neuron that drives the positive feedback amplification of correlated activity is a key factor underlying the destabilization of synchrony in smaller networks. Spontaneous neural activity plays a critical role in cortical information processing, and our work constructively clarifies an aspect of the structural basis behind this.

Quantitative comparison of cancer and normal cell adhesion using organosilane monolayer templates: an experimental study on the anti-adhesion effect of green-tea catechins.

The main constituent of green tea, (-)-Epigallocatechin-3-O-gallate (EGCG), is known to have cancer-specific chemopreventive effects. In the present work, we investigated how EGCG suppresses cell adhesion by comparing the adhesion of human pancreatic cancer cells (AsPC-1 and BxPC-3) and their counterpart, normal human embryonic pancreas-derived cells (1C3D3), in catechin-containing media using organosilane monolayer templates (OMTs). The purpose of this work is (1) to evaluate the quantitativeness in the measurement of cell adhesion with the OMT and (2) to show how green-tea catechins suppress cell adhesion in a cancer-specific manner. For the first purpose, the adhesion of cancer and normal cells was compared using the OMT. The cell adhesion in different type of catechins such as EGCG, (-)-Epicatechin-3-O-gallate (ECG) and (-)-Epicatechin (EC) was also evaluated. The measurements revealed that the anti-adhesion effect of green-tea catechins is cancer-specific, and the order is EGCG≫ECG>EC. The results agree well with the data reported to date, showing the quantitativeness of the new method. For the second purpose, the contact area of cells on the OMT was measured by reflection interference contrast microscopy. The cell-OMT contact area of cancer cells decreases with increasing EGCG concentration, whereas that of normal cells remains constant. The results reveal a twofold action of EGCG on cancer cell adhesion-suppressing cell attachment to a candidate adhesion site and decreasing the contact area of the cells-and validates the use of OMT as a tool for screening cancer cell adhesion.

Quantitative Evaluation of Cancer Cell Adhesion to Self-Assembled Monolayer-Patterned Substrates by Reflection Interference Contrast Microscopy.

Adhesion of cancer cells with different metastatic potential and anticancer drug resistance has been quantitatively evaluated by using self-assembled monolayer (SAM)-patterned substrates and reflection interference contrast microscopy (RICM). Cell-adhesive SAM spots with optimized diameter could prevent cell-cell adhesion and thus allowed the systematic evaluation of statistically reliable numbers of contact area between single cancer cells and substrates by RICM. The statistical image analysis revealed that highly metastatic mouse melanoma cells showed larger contact area than lowly metastatic cells. We also found that both cancer cell types exhibited distinct transition from the "strong" to "weak" adhesion states with increase in the concentration of (-)-epigallocatechin gallate (EGCG), which is known to exhibit cancer preventive activity. Mathematical analysis of the adhesion transition revealed that adhesion of the highly metastatic mouse melanoma cells showed more EGCG tolerance than that of lowly metastatic cells. Moreover, time-lapse RICM observation revealed that EGCG weakened cancer cell adhesion in a stepwise manner, probably via focal adhesion complex. These results clearly indicate that contact area can be used as a quantitative measure for the determination of cancer phenotypes and their drug resistance, which will provide physical insights into the mechanism of cancer metastasis and cancer prevention.

Neuraminidase-Dependent Degradation of Polysialic Acid Is Required for the Lamination of Newly Generated Neurons.

Hippocampal granule cells (GCs) are generated throughout the lifetime and are properly incorporated into the innermost region of the granule cell layer (GCL). Hypotheses for the well-regulated lamination of newly generated GCs suggest that polysialic acid (PSA) is present on the GC surface to modulate GC-to-GC interactions, regulating the process of GC migration; however, direct evidence of this involvement is lacking. We show that PSA facilitates the migration of newly generated GCs and that the activity of N-acetyl-α-neuraminidase 1 (NEU1, sialidase 1) cleaves PSA from immature GCs, terminating their migration in the innermost GCL. Developing a migration assay of immature GCs in vitro, we found that the pharmacological depletion of PSA prevents the migration of GCs, whereas the inhibition of PSA degradation with a neuraminidase inhibitor accelerates this migration. We found that NEU1 is highly expressed in immature GCs. The knockdown of NEU1 in newly generated GCs in vivo increased PSA presence on these cells, and attenuated the proper termination of GC migration in the innermost GCL. In conclusion, this study identifies a novel mechanism that underlies the proper lamination of newly generated GCs through the modulation of PSA presence by neuronal NEU1.

Photopatterning Proteins and Cells in Aqueous Environment Using TiO2 Photocatalysis.

Organic contaminants adsorbed on the surface of titanium dioxide (TiO2) can be decomposed by photocatalysis under ultraviolet (UV) light. Here we describe a novel protocol employing the TiO2 photocatalysis to locally alter cell affinity of the substrate surface. For this experiment, a thin TiO2 film was sputter-coated on a glass coverslip, and the TiO2 surface was subsequently modified with an organosilane monolayer derived from octadecyltrichlorosilane (OTS), which inhibits cell adhesion. The sample was immersed in a cell culture medium, and focused UV light was irradiated to an octagonal region. When a neuronal cell line PC12 cells were plated on the sample, cells adhered only on the UV-irradiated area. We further show that this surface modification can also be performed in situ, i.e., even when cells are growing on the substrate. Proper modification of the surface required an extracellular matrix protein collagen to be present in the medium at the time of UV irradiation. The technique presented here can potentially be employed in patterning multiple cell types for constructing coculture systems or to arbitrarily manipulate cells under culture.

Fluorescence Polarization Switching from a Single Silicon Vacancy Colour Centre in Diamond.

Single-photon emitters with stable and uniform photoluminescence properties are important for quantum technology. However, in many cases, colour centres in diamond exhibit spectral diffusion and photoluminescence intensity fluctuation. It is therefore essential to investigate the dynamics of colour centres at the single defect level in order to enable the on-demand manipulation and improved applications in quantum technology. Here we report the polarization switching, intensity jumps and spectral shifting observed on a negatively charged single silicon-vacancy colour centre in diamond. The observed phenomena elucidate the single emitter dynamics induced by photoionization of nearby electron donors in the diamond.

Investigation of the silicon vacancy color center for quantum key distribution.

Single photon sources (SPS) are crucial for quantum key distribution. Here we demonstrate a stable triggered SPS at 738 nm with linewidth less than 5 nm at room temperature based on a negatively charged single silicon vacancy color center. Thanks to the short photon duration of about 1.3-1.7 ns, by using high repetition pulsed excitation at 30 MHz, the triggered single photon source generates 16.6 kcounts/s. And we discuss the feasibility of this triggered SPS in the application of quantum key distribution.

In situ modification of cell-culture scaffolds by photocatalytic decomposition of organosilane monolayers.

We demonstrate a novel application of TiO2 photocatalysis for modifying the cell affinity of a scaffold surface in a cell-culture environment. An as-deposited octadecyltrichlorosilane self-assembled monolayer (OTS SAM) on TiO2 was found to be hydrophobic and stably adsorbed serum albumins that blocked subsequent adsorption of other proteins and cells. Upon irradiation of ultraviolet (UV) light, OTS molecules were decomposed and became permissive to the adhesion of PC12 cells via adsorption of an extracellular matrix protein, collagen. Optimal UV dose was 200 J cm(-2) for OTS SAM on TiO2. The amount of collagen adsorption decreased when excessive UV light was irradiated, most likely due to the surface being too hydrophilic to support its adsorption. This UV-induced modification required TiO2 to be present under the SAM and hence is a result of TiO2 photocatalysis. The UV irradiation for surface modification can be performed before cell plating or during cell culture. We also demonstrate that poly(ethylene glycol) SAM can also be patterned with this method, indicating that it is applicable to both hydrophobic and hydrophilic SAMs. This method provides a unique tool for fabricating cell microarrays and studying dynamical properties of living cells.

Improving zero-mode waveguide structure for enhancing signal-to-noise ratio of real-time single-molecule fluorescence imaging: a computational study.

We investigated the signal-to-noise ratio (S/N) of real-time single-molecule fluorescence imaging (SMFI) using zero-mode waveguides (ZMWs). The excitation light and the fluorescence propagating from a molecule in the ZMW were analyzed by computational optics simulation. The dependence of the S/N on the ZMW structure was investigated with the diameter and etching depth as the simulation parameters. We found that the SMFI using a conventional ZMW was near the critical level for detecting binding and dissociation events. We show that etching the glass surface of the ZMW by 60 nm enhances the S/N six times the conventional nonetched ZMWs. The enhanced S/N improves the temporal resolution of the SMFI at physiological concentrations.

SPECT/CT imaging in ulnocarpal impaction syndrome.

We report a 42-year-old man who presented with ulnar-sided wrist pain. MR image revealed bone marrow edema in the lunate bone. Ulnocarpal impaction syndrome was diagnosed by MRI and arthrography. Bone scintigraphy showed increased uptake in the wrist. SPECT/CT with 2- and 3-dimensional reconstruction images clearly demonstrated abnormal accumulation on the ulnar side of the lunate bone.