Organotypic slice culture of embryonic brain tissue.
Organotypic slice culture of embryonic brain tissue.
复制标题
DOI：
10.1101/pdb.prot4914
复制
发表时间：
2007-12-01
期刊：
影响因子：
--
通讯作者：
Hevner, Robert F
中科院分区：
文献类型：
--
作者：
Daza, Ray A M;Englund, Chris;Hevner, Robert F
本方案描述了如何解剖、组装和培养E11.5至E18.5(天)的小鼠胚胎(E)脑组织，用于器官型切片培养。这些制剂可用于各种分析和研究，包括不同脑区的共培养、细胞迁移分析、轴突引导分析和DNA电穿孔实验。在电穿孔过程中，将电流施加到脑片特定靶区的表面，以便在质膜上打开孔，并引入编码DNA的质粒。浮动的膜上切片结构有助于保持脑切片的结构完整性，同时保持容易的实验通道和最佳的生存能力。可以在活切片中监测实验(例如，使用共聚焦成像)，并且可以使用在实验结束时固定和冷冻的切片来完成进一步的研究。胚胎大脑或脊椎组织的任何区域都可以用于这一方案。
INTRODUCTIONThis protocol describes how to dissect, assemble, and cultivate mouse embryonic (E) brain tissue from age E11.5 to E18.5 (days) for organotypic slice culture. These preparations can be used for a variety of assays and studies including coculture of different brain regions, cell migration assays, axon guidance assays, and DNA electroporation experiments. During electroporation, an electric current is applied to the surface of a specific target area of the brain slice in order to open holes in the plasma membrane and introduce a plasmid of coding DNA. The floating slice-on-membrane construct helps to preserve the structural integrity of the brain slices, while maintaining easy experimental access and optimal viability. Experiments can be monitored in living slices (e.g., with confocal imaging), and further studies can be completed using slices that have been fixed and cryosectioned at the end of the experiment. Any region of embryonic brain or spinal tissue can be used in this protocol.