Center for Cell and Developmental Biology, State Key Laboratory for Agrobiotechnology, School of Life Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
Methods Mol Biol. 2020;2177:59-67. doi: 10.1007/978-1-0716-0767-1_6.
The plant trans-Golgi network (TGN) is a multifunctional organelle derived from the Golgi. It consists of tubulovesicular compartments scattered in the cytosol. They produce secretory vesicles delivering proteins and polysaccharides to the cell wall. They also serve as early endosomal compartments, receiving endocytic cargos from the plasma membrane. This versatility is thought to originate from functional variations among individual TGN compartments. Correlative light and electron microscopy (CLEM) combines the imaging capability of light microscopy and electron microscopy (EM) to determine the location of macromolecules in EM images in the cellular context. It is possible to identify organelles associated with specific fluorescent markers and examine their membrane architectures at nanometer-level resolutions using CLEM. In this chapter, we will explain the CLEM method that our lab uses to investigate functional and structural heterogeneity among individual TGN compartments in plant cells.
植物反式高尔基体网络（TGN）是一种源自高尔基体的多功能细胞器。它由分散在细胞质中的管状泡状隔室组成。它们产生分泌小泡，将蛋白质和多糖输送到细胞壁。它们还充当早期内体隔室，从质膜接收内吞货物。这种多功能性被认为源于单个 TGN 隔室之间的功能变化。相关的光镜和电子显微镜（CLEM）将光镜和电子显微镜（EM）的成像能力结合起来，以确定细胞环境中 EM 图像中大分子的位置。使用 CLEM，可以识别与特定荧光标记物相关的细胞器，并以纳米级分辨率检查它们的膜结构。在本章中，我们将解释我们实验室使用的 CLEM 方法，以研究植物细胞中单个 TGN 隔室之间的功能和结构异质性。
