Jeon Noo Li
Department of Biomedical Engineering, University of California, Irvine, CA, USA.
J Vis Exp. 2007(9):411. doi: 10.3791/411. Epub 2007 Nov 1.
This video describes the fabrication and use of a microfluidic device to culture central nervous system (CNS) neurons. This device is compatible with live-cell optical microscopy (DIC and phase contrast), as well as confocal and two photon microscopy approaches. This method uses precision-molded polymer parts to create miniature multi-compartment cell culture with fluidic isolation. The compartments are made of tiny channels with dimensions that are large enough to culture neurons in well-controlled fluidic microenvironments. Neurons can be cultured for 2-3 weeks within the device, after which they can be fixed and stained for immunocytochemistry. Axonal and somal compartments can be maintained fluidically isolated from each other by using a small hydrostatic pressure difference; this feature can be used to localize soluble insults to one compartment for up to 20 h after each medium change. Fluidic isolation enables collection of pure axonal fraction and biochemical analysis by PCR. The microfluidic device provides a highly adaptable platform for neuroscience research and may find applications in modeling CNS injury and neurodegeneration.
本视频介绍了一种用于培养中枢神经系统（CNS）神经元的微流控装置的制造和使用方法。该装置适用于活细胞光学显微镜（微分干涉对比显微镜和相差显微镜），以及共聚焦显微镜和双光子显微镜方法。此方法利用精密模制的聚合物部件来创建具有流体隔离功能的微型多隔室细胞培养环境。隔室由微小通道构成，其尺寸大到足以在良好控制的流体微环境中培养神经元。神经元可在该装置内培养2至3周，之后可进行固定并用于免疫细胞化学染色。通过利用小的静水压力差，轴突隔室和胞体隔室可在流体上保持相互隔离；每次更换培养基后，这一特性可用于将可溶性损伤定位在一个隔室内长达20小时。流体隔离能够收集纯轴突部分并通过聚合酶链反应进行生化分析。该微流控装置为神经科学研究提供了一个高度适应性强的平台，可能在中枢神经系统损伤和神经退行性变建模方面找到应用。
