1] Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Kowloon, Hong Kong SAR [2].
Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Kowloon, Hong Kong SAR.
Sci Rep. 2014 Apr 24;4:4784. doi: 10.1038/srep04784.
Recreation of neural network in vitro with designed topology is a valuable tool to decipher how neurons behave when interacting in hierarchical networks. In this study, we developed a simple and effective platform to pattern primary neurons in array formats for interrogation of neural circuitry with single cell resolution. Unlike many surface-chemistry-based patterning methods, our NeuroArray technique is specially designed to accommodate neuron's polarized morphologies to make regular arrays of cells without restricting their neurite outgrowth, and thus allows formation of freely designed, well-connected, and spontaneously active neural network. The NeuroArray device was based on a stencil design fabricated using a novel sacrificial-layer-protected PDMS molding method that enables production of through-structures in a thin layer of PDMS with feature sizes as small as 3 µm. Using the NeuroArray along with calcium imaging, we have successfully demonstrated large-scale tracking and recording of neuronal activities, and used such data to characterize the spiking dynamics and transmission within a diode-like neural network. Essentially, the NeuroArray is a universal patterning platform designed for, but not limited to neuron cells. With little adaption, it can be readily interfaced with other interrogation modalities for high-throughput drug testing, and for building neuron culture based live computational devices.
利用设计的拓扑结构在体外重建神经网络是破译神经元在分层网络中相互作用时行为的一种有价值的工具。在这项研究中，我们开发了一种简单有效的平台，用于以阵列格式对原代神经元进行图案化，以便以单细胞分辨率研究神经回路。与许多基于表面化学的图案化方法不同，我们的 NeuroArray 技术专门设计用于适应神经元的极化形态，以便在不限制其突起生长的情况下形成规则的细胞阵列，从而允许形成自由设计、良好连接和自发活跃的神经网络。NeuroArray 设备基于使用新型牺牲层保护 PDMS 成型方法制造的模板设计，该方法能够在薄的 PDMS 层中生产尺寸小至 3 µm 的通孔结构。使用 NeuroArray 结合钙成像，我们已经成功地演示了大规模跟踪和记录神经元活动，并使用这些数据来表征二极管样神经网络中的尖峰动力学和传输。从本质上讲，NeuroArray 是一个通用的图案化平台，专为神经元细胞设计，但不限于神经元细胞。稍加修改，它就可以很容易地与其他检测模式接口，用于高通量药物测试，并用于构建基于神经元培养的活体计算设备。
