Paranjape Smita R, Nagendran Tharkika, Poole Valerie, Harris Joseph, Taylor Anne Marion
UNC Neuroscience Center; UNC/NC State Joint Department of Biomedical Engineering.
Xona Microfluidics, LLC.
J Vis Exp. 2019 May 3(147). doi: 10.3791/59250.
Use of microfluidic devices to compartmentalize cultured neurons has become a standard method in neuroscience. This protocol shows how to use a pre-assembled multi-compartment chip made in a cyclic olefin copolymer (COC) to compartmentalize neurons differentiated from human stem cells. The footprint of these COC chips are the same as a standard microscope slide and are equally compatible with high resolution microscopy. Neurons are differentiated from human neural stem cells (NSCs) into glutamatergic neurons within the chip and maintained for 5 weeks, allowing sufficient time for these neurons to develop synapses and dendritic spines. Further, we demonstrate multiple common experimental procedures using these multi-compartment chips, including viral labeling, establishing microenvironments, axotomy, and immunocytochemistry.
使用微流控装置对培养的神经元进行分隔已成为神经科学中的一种标准方法。本方案展示了如何使用由环烯烃共聚物（COC）制成的预组装多隔室芯片对源自人类干细胞的分化神经元进行分隔。这些COC芯片的尺寸与标准显微镜载玻片相同，并且同样适用于高分辨率显微镜检查。神经元在芯片内从人类神经干细胞（NSCs）分化为谷氨酸能神经元，并维持5周，这为这些神经元发育突触和树突棘留出了足够的时间。此外，我们展示了使用这些多隔室芯片的多个常见实验程序，包括病毒标记、建立微环境、轴突切断和免疫细胞化学。
