A 3D neuronal network read-out interface with high recording performance using a neuronal cluster patterning on a microelectrode array.

In recent years, in vitro three-dimensional (3D) neuronal network models utilizing extracellular matrices have been advancing. To understand the network activity from these models, attempts have been made to measure activity in multiple regions simultaneously using a microelectrode array (MEA). Although there hve been many attempts to measure the activity of 3D networks using 2-dimensional (2D) MEAs, the physical coupling between the 3D network and the microelectrodes was not stable and needed to be improved. In this study, we proposed a neuronal cluster interface that improves the active channel ratio of commercial 2D MEAs, enabling reliable measurement of 3D network activity. To achieve this, neuronal clusters, which consist of a small number of neurons, were patterned on microelectrodes and used as mediators to transmit the signal between the 3D network and the microelectrodes. We confirmed that the patterned neuronal clusters enhanced the active channel ratio and SNR(signal-to-noise-ratio) about 3D network recording and stimulation for a month. Our interface was able to functionally connect with 3D networks and measure the 3D network activity without significant alternation of activity characteristics. Finally, we demonstrated that our interface can be used to analyze the differences in the dynamics of 3D and 2D networks and to construct the 3D clustered network. This method is expected to be useful for studying the functional activity of various 3D neuronal network models, offering broad applications for the use of these models.

Enhancement of Thermoplasmonic Neural Modulation Using a Gold Nanorod-Immobilized Polydopamine Film.

Photothermal neural activity inhibition has emerged as a minimally invasive neuromodulation technology with submillimeter precision. One of the techniques involves the utilization of plasmonic gold nanoparticles (AuNPs) to modulate neural activity by photothermal effects ("thermoplasmonics"). A surface modification technique is often required to integrate AuNPs onto the neural interface. Here, polydopamine (pDA), a multifunctional adhesive polymer with a wide light absorption spectrum, is introduced both as a primer layer for the immobilization of gold nanorods (GNRs) on the neural interface and as an additional photothermal agent by absorbing near-infrared red (NIR) lights for more efficient photothermal effects. First, the optical and photothermal properties of pDA as well as the characteristics of GNRs attached onto the pDA film are investigated for the optimized photothermal neural interface. Due to the covalent bonding between GNR surfaces and pDA, GNRs immobilized on pDA showed strong attachment onto the surface, yielding a more stable photothermal platform. Lastly, when photothermal neural stimulation was applied to the primary rat hippocampal neurons, the substrate with GNRs immobilized on the pDA film allowed more laser power-efficient photothermal neuromodulation as well as photothermal cell death. This study suggests the feasibility of using pDA as a surface modification material for developing a photothermal platform for the inhibition of neural activities.

Development of the micro-patterned 3D neuronal-hydrogel model using soft-lithography for study a 3D neural network on a microelectrode array.

In vitro patterned neuronal models have been studied as one of the strategies to investigate the relationship between structural connectivity and functional activity of neural network. Despite the importance of three-dimensional (3D) cell models, most of these studies have been performed on two-dimensional models. In this study, we present a technique to construct the micro-pattern to 3D neuronal-hydrogel model using a micromolding in capillaries (MIMIC) technique on microelectrode array (MEA). Our technique was suitable to prevent the deformation of micro-patterned collagen model against the neuronal contracted tension during the network formation. The relationship between the growth directions of glial cells and micro-pattern direction was investigated. Lastly, we confirmed that our 3D model had synchronized activity among neurons in 3D. This model is expected to be used as a tool to study the relationship between structural connectivity and functional activity in the 3D environment.

Label-free monitoring of 3D cortical neuronal growth in vitro using optical diffraction tomography.

The highly complex central nervous systems of mammals are often studied using three-dimensional (3D) in vitro primary neuronal cultures. A coupled confocal microscopy and immunofluorescence labeling are widely utilized for visualizing the 3D structures of neurons. However, this requires fixation of the neurons and is not suitable for monitoring an identical sample at multiple time points. Thus, we propose a label-free monitoring method for 3D neuronal growth based on refractive index tomograms obtained by optical diffraction tomography. The 3D morphology of the neurons was clearly visualized, and the developmental processes of neurite outgrowth in 3D spaces were analyzed for individual neurons.