Subcellular Stucture in Glial Networks by TEM

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，使用高压冷冻。培养的细胞在蓝宝石盘上生长，在那里它们可以通过光学显微镜进行评估。在适当的时间，将圆片在液氮温度下高压冷冻以保存超微结构。然后将标本用锇固定剂冷冻替代，然后包埋在塑料中，切片并染色。实验条件正在调查，是适合结构研究。我们的目的是在300千伏的能量过滤透射电子显微镜中使用电子断层扫描成像的神经胶质细胞在三维空间中的相互作用区。

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 was used. Cultured cells were grown on sapphire discs where they could be assessed by optical microscopy. At the appropriate time, the discs were high-pressure frozen at liquid nitrogen temperature to preserve ultrastructure. Specimens were then freeze-substituted with osmium fixative before being embedded in plastic, sectioned and stained. Experimental conditions are being investigated that are suitable for structural studies. The aim is to image interaction zones of the glial cells in three dimensions by using electron tomography in a 300 kV energy-filtering transmission electron microscope.