Schürmann Matthias, Shepheard Norman, Frese Natalie, Geishendorf Kevin, Sudhoff Holger, Gölzhäuser Armin, Rückert Ulrich, Kaltschmidt Christian, Kaltschmidt Barbara, Thomas Andy
Cell Biology, Bielefeld University, Bielefeld, Germany.
Department of Otolaryngology, Head and Neck Surgery, Klinikum Bielefeld, Bielefeld, Germany.
PLoS One. 2018 Feb 23;13(2):e0192647. doi: 10.1371/journal.pone.0192647. eCollection 2018.
In this manuscript, we first reveal a simple ultra violet laser lithographic method to design and produce plain tailored multielectrode arrays. Secondly, we use the same lithographic setup for surface patterning to enable controlled attachment of primary neuronal cells and help neurite guidance. For multielectrode array production, we used flat borosilicate glass directly structured with the laser lithography system. The multi layered electrode system consists of a layer of titanium coated with a layer of di-titanium nitride. Finally, these electrodes are covered with silicon nitride for insulation. The quality of the custom made multielectrode arrays was investigated by light microscopy, electron microscopy and X-ray diffraction. The performance was verified by the detection of action potentials of primary neurons. The electrical noise of the custom-made MEA was equal to commercially available multielectrode arrays. Additionally, we demonstrated that structured coating with poly lysine, obtained with the aid of the same lithographic system, could be used to attach and guide neurons to designed structures. The process of neuron attachment and neurite guidance was investigated by light microscopy and charged particle microscopy. Importantly, the utilization of the same lithographic system for MEA fabrication and poly lysine structuring will make it easy to align the architecture of the neuronal network to the arrangement of the MEA electrode.. In future studies, this will lead to multielectrode arrays, which are able to specifically attach neuronal cell bodies to their chemically defined electrodes and guide their neurites, gaining a controlled connectivity in the neuronal network. This type of multielectrode array would be able to precisely assign a signal to a certain neuron resulting in an efficient way for analyzing the maturation of the neuronal connectivity in small neuronal networks.
在本论文中，我们首先揭示了一种简单的紫外激光光刻方法，用于设计和制造普通定制的多电极阵列。其次，我们使用相同的光刻装置进行表面图案化，以实现原代神经元细胞的可控附着并辅助神经突导向。对于多电极阵列的制造，我们使用激光光刻系统直接对平面硼硅酸盐玻璃进行结构化处理。多层电极系统由一层钛和一层二氮化钛涂层组成。最后，这些电极覆盖有氮化硅用于绝缘。通过光学显微镜、电子显微镜和X射线衍射对定制的多电极阵列的质量进行了研究。通过检测原代神经元的动作电位验证了其性能。定制的MEA的电噪声与市售多电极阵列相当。此外，我们证明了借助相同的光刻系统获得的聚赖氨酸结构化涂层可用于将神经元附着并引导至设计结构。通过光学显微镜和带电粒子显微镜研究了神经元附着和神经突导向的过程。重要的是，在MEA制造和聚赖氨酸结构化中使用相同的光刻系统将便于使神经网络的结构与MEA电极的排列对齐。在未来的研究中，这将导致多电极阵列能够将神经元细胞体特异性地附着到其化学定义的电极上并引导其神经突，从而在神经网络中获得可控的连接性。这种类型的多电极阵列将能够精确地将信号分配给特定的神经元，从而以一种有效的方式分析小型神经网络中神经元连接的成熟情况。
