用于研究光呼吸作用的光学显微镜、透射电子显微镜及免疫组织化学实验方案

Light Microscopy, Transmission Electron Microscopy, and Immunohistochemistry Protocols for Studying Photorespiration.

作者信息

Khoshravesh Roxana, Lundsgaard-Nielsen Vanessa, Sultmanis Stefanie, Sage Tammy L

机构信息

Department of Ecology and Evolutionary Biology, University of Toronto, 25 Willcocks St., Toronto, ON, Canada, M5S 3B2.

出版信息

Methods Mol Biol. 2017;1653:243-270. doi: 10.1007/978-1-4939-7225-8_17.

DOI:10.1007/978-1-4939-7225-8_17

PMID:28822138

Abstract

High-resolution images obtained from plant tissues processed for light microscopy, transmission electron microscopy, and immunohistochemistry have provided crucial links between plant subcellular structure and physiology during photorespiration as well as the impact of photorespiration on plant evolution and development. This chapter presents established protocols to guide researchers in the preparation of plant tissues for high-resolution imaging with a light and transmission electron microscope and detection of proteins using immunohistochemistry. Discussion of concepts and theory behind each step in the process from tissue preservation to staining of resin-embedded tissues is included to enhance the understanding of all steps in the procedure. We also include a brief protocol for quantification of cellular parameters from high-resolution images to help researchers rigorously test hypotheses.

摘要

从经过光镜、透射电子显微镜和免疫组织化学处理的植物组织中获得的高分辨率图像，为光呼吸过程中植物亚细胞结构与生理之间的关键联系以及光呼吸对植物进化和发育的影响提供了重要线索。本章介绍了既定的方案，以指导研究人员制备用于光镜和透射电子显微镜高分辨率成像的植物组织，以及使用免疫组织化学检测蛋白质。文中包括了从组织保存到树脂包埋组织染色过程中每一步背后的概念和理论讨论，以增进对该过程所有步骤的理解。我们还包括了一个从高分辨率图像定量细胞参数的简要方案，以帮助研究人员严格检验假设。