Analysis of microglial migration by a micropipette assay
Analysis of microglial migration by a micropipette assay
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通过微量移液器测定分析小胶质细胞迁移
DOI：
10.1038/nprot.2014.015
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发表时间：
2014-02-01
期刊：
影响因子：
14.8
通讯作者：
Duan, Shumin
中科院分区：
文献类型：
--
作者：
Wu, Hang-jun;Liu, Yi-jun;Duan, Shumin
小胶质细胞在维持脑内环境稳定方面具有重要作用，并且它们与多种脑疾病有关。目前有兴趣研究小胶质细胞迁移，导致细胞积聚在损伤的焦点部位。在这里，我们描述了一个协议，快速触发和监测小胶质细胞迁移使用微量移液管测定。该方案是使用小胶质细胞的轴突转向测定的适应。从微量移液器尖端释放的化学引诱剂产生诱导强大的小胶质细胞迁移的趋化梯度。结合显微镜成像，该测定允许同时记录细胞运动和亚细胞区室运输，沿着定量分析。试验的实际处理时间总计约为2-3小时。该方案简单，廉价，易于建立，可用于检查多种细胞类型的细胞迁移，包括具有广泛化学信号的癌细胞。
Microglial cells have important roles in maintaining brain homeostasis, and they are implicated in multiple brain diseases. There is currently interest in investigating microglial migration that results in cell accumulation at focal sites of injury. Here we describe a protocol for rapidly triggering and monitoring microglial migration by using a micropipette assay. This protocol is an adaptation of the axon turning assay using microglial cells. Chemoattractants released from the micropipette tip produce a chemotactic gradient that induces robust microglial migration. In combination with microscopic imaging, this assay allows simultaneous recording of cell movement and subcellular compartment trafficking, along with quantitative analysis. The actual handling time for the assay takes similar to 2-3 h in total. The protocol is simple, inexpensive and convenient to set up, and it can be adopted to examine cell migration in multiple cell types, including cancer cells with a wide range of chemical signals.