Subcellular Stucture in Glial Networks by Transmission E

本研究的目的是在超分子水平上研究神经胶质细胞线粒体和内质网相邻区域的超微结构特征。这两种类型的细胞器之间的相互作用与钙信号有关，通过钙信号，神经胶质细胞在中枢神经系统发育和突触活动期间相互交流。为了将神经胶质网络保持在其生理状态，已经应用了高压冷冻。培养的细胞首先在特殊涂覆的金样品载体上生长，然后在液氮温度下在>2000个大气压的压力下冷冻，以避免冰晶损伤。然后在塑料包埋、切片和染色之前，用锇固定剂冷冻替代标本。神经胶质细胞中的感兴趣区域正在使用数字能量过滤透射电子显微镜中的电子断层扫描进行三维成像。

The aim of this project is to study ultrastructural features at the supramolecular scale in glial cells in the zones where mitochondria and endoplasmic reticulum adjoin. Interactions between these two types of organelle are associated with calcium signaling, through which glial cells communicate with each other during central nervous system development and synaptic activity. To preserve glial networks in their physiological state, high-pressure freezing has been applied. Cultured cells are first grown on specially coated gold specimen carriers, which are then frozen at a pressure of >2000 atmospheres at liquid nitrogen temperature to avoid ice crystal damage. Specimens are then freeze-substituted with osmium fixative before plastic embedding, sectioning and staining. The zones of interest in the glial cells are being imaged in three dimensions using electron tomography in a digital energy-filtering transmission electron microscope.