高压冷冻和冷冻置换用于透射电子显微镜成像和免疫金标记。

High-Pressure Freezing and Freeze Substitution for Transmission Electron Microscopy Imaging and Immunogold-Labeling.

机构信息

Department of Botany, University of Wisconsin-Madison, Madison, WI, USA.

Department of Genetics, University of Wisconsin-Madison, Madison, WI, USA.

出版信息

Methods Mol Biol. 2021;2200:337-347. doi: 10.1007/978-1-0716-0880-7_16.

DOI:10.1007/978-1-0716-0880-7_16

PMID:33175386

Abstract

Electron microscopy enables the unbiased imaging of organelles and cellular structures at nano-meter scale resolution. The combination of cryofixation/freeze-substitution methods with other imaging techniques such as correlative light and electron microscopy (CLEM), electron tomography (ET), and immunogold-labeling provides unique opportunities to understand structural changes associated with cellular processes. This chapter presents the main steps in the preparation of Arabidopsis thaliana roots, cotyledons, anthers, and developing seeds by high-pressure freezing and freeze-substitution for structural analysis and immunogold-labeling using transmission electron microscopy.

摘要

电子显微镜能够以纳米级分辨率对细胞器和细胞结构进行无偏成像。冷冻固定/冷冻置换方法与其他成像技术（如相关光和电子显微镜（CLEM）、电子断层扫描（ET）和免疫金标记）的结合，为理解与细胞过程相关的结构变化提供了独特的机会。本章介绍了通过高压冷冻和冷冻置换制备拟南芥根、子叶、花粉和发育种子用于结构分析和免疫金标记的透射电子显微镜的主要步骤。