Processing plant tissues for ultrastructural study

Research output: Chapter in Book/Conference paperChapter

4 Citations (Scopus)

Abstract

This chapter briefl y describes conventional and microwave-assisted chemical fixation methods, as well as cryo-specimen preparation techniques for studying the cellular and organelle ultrastructure of plant tissues under transmission electron microscopy. The general methods and procedures for the plant specimen preparation (including fixation, dehydration, resin infi ltration, and embedding) are similar to those for animal tissues. However, certain special characteristic features of plant tissues such as thick cellulosic cell wall, waxy substance in the cuticle, large amount of gases in the intercellular spaces, and the presence of vacuoles have created fixation and resin filtration diffi culties. Specifi c modifi cations of the protocols used for animal tissues are therefore required, such as the application of vacuum during the initial fixation and resin infi ltration stage to remove gases from the tissues and resin. Microwave-assisted procedure can reduce specimen preparation time, but both conventional and microwave-assisted chemical fixation procedures produce artifacts. Cryo-specimen preparation involves with high-pressure freezing and freeze-substitution can minimize artifact formation, but their application to highly vacuolated, thick-walled plant cells is limited. © Springer Science+Business Media New York 2014.
Original languageEnglish
Title of host publicationElectron Microscopy: Methods and Protocols
EditorsJohn Kuo
Place of PublicationNew York
PublisherHumana Press
Pages39-55
Volume1117
Edition3
ISBN (Print)9781627037754
DOIs
Publication statusPublished - 2014

Publication series

NameMethods in Molecular Biology

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