This second edition provides new and updated methods detailing techniques and state-of-the-art approaches on the structure and function of plant endoplasmic reticulum (ER). Chapters guide readers through modern microscopy techniques, software protocols, purification, and analysis of ER membrane structure. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.