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通过印刷石墨烯微电极对神经细胞进行分钟级敏感的实时监测。

Minute-sensitive real-time monitoring of neural cells through printed graphene microelectrodes.

机构信息

Department of Mechanical Engineering, Iowa State University, Ames, IA, 50011, USA.

Department of Mechanical Engineering, Iowa State University, Ames, IA, 50011, USA.

出版信息

Biosens Bioelectron. 2022 Aug 15;210:114284. doi: 10.1016/j.bios.2022.114284. Epub 2022 Apr 17.

Abstract

Real-time and high-throughput cytometric monitoring of neural cells exposed to injury mechanisms is invaluable for in-vitro studies. Electrical impedance spectroscopy via microelectrode arrays is a label-free technique for monitoring of neural growth and their detachment upon death. In this method, the interface material plays a vital role to provide desirable attachment cues for the cell network. Thus, here we demonstrate the electrohydrodynamic patterning of aqueous graphene for microelectrode fabrication. We investigated whether the wrinkled surface morphology of the electrodes fabricated by this deposition method expands their electroactive surface area and thus enables a rapid response time. The nano-scale quality of the graphene lattice is characterized by Raman spectroscopy and Transmittance electron microscopy. N27 rat dopaminergic neural cells were cultured on the chips and the surface morphology of the microelectrodes during cellular growth was investigated by Scanning electrode spectroscopy. Attachment of the neural population on the graphene microelectrodes was parametrized and the change in the impedance spectrum of this cell population was quantified at 10 Hz to 10 kHz frequencies along with the change in TUBB3 gene expression. The viability test of the cell population on the biosensor demonstrated no significant difference in comparison to the control, and a cell density of 2289 cell/mm was achieved. As a proof of concept, the confluent N27 cell population was exposed to UV and its cytotoxic impact on neural detachment and lift-off was monitored. The multiplexed detection of cellular activity was reported with a temporal resolution of one minute.

摘要

实时高通量细胞仪监测神经细胞暴露于损伤机制对于体外研究是非常宝贵的。通过微电极阵列的介电电泳谱是监测神经生长及其死亡时脱落的无标记技术。在这种方法中,界面材料起着至关重要的作用,为细胞网络提供理想的附着线索。因此,我们在此展示了用于微电极制造的水石墨烯的电动图案化。我们研究了这种沉积方法制造的电极的褶皱表面形态是否会扩大其电活性表面积,从而实现快速响应时间。石墨烯晶格的纳米级质量通过拉曼光谱和透射电子显微镜进行表征。N27 大鼠多巴胺能神经细胞在芯片上培养,并通过扫描电极光谱研究细胞生长过程中微电极的表面形态。对神经群体在石墨烯微电极上的附着进行参数化,并在 10 Hz 至 10 kHz 频率下定量测量该细胞群体的阻抗谱变化,同时测量 TUBB3 基因表达的变化。与对照组相比,细胞群体在生物传感器上的活力测试没有明显差异,并且实现了 2289 个细胞/mm 的细胞密度。作为概念验证,将密集的 N27 细胞群体暴露于 UV 下,并监测其对神经细胞脱落和脱离的细胞毒性影响。细胞活性的多重检测报告了一分钟的时间分辨率。

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