Processing Plant Tissues for Ultrastructural Study
By: John Kuo2
| Abstract |
|
|
This chapter describes conventional chemical fixation methods and 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, infiltration, and embedding) and the composition of fixatives, buffers, dehydration solvent,
and embedding media are similar to those for animal tissues. However, certain special characteristic features of plant tissues,
such as a thick cellulosic cell wall, waxy substance in the cuticle, large amount of gases in the intercellular spaces, the
presence of vacuoles, have created fixation and resin filtration difficulties and, therefore, special modifications of the
protocols used for animal tissues are required. The addition of chemicals such as caffeine in fixative can stabilize the phenol
in the vacuole; however, the rupture of vacuole caused by the fixative still cannot be controlled, particularly for plants
with highly vacuolated cells. The application of vacuum infiltration during the initial fixation stage to remove gases from
the tissues is described. Additional vacuum infiltration during resin infiltration procedure to improve the efficiency of
resin penetration is implemented.
Affiliation(s): (2) Centre For Microscopy and Microanalysis, The University of Western Australia, Crawley, Western Australia, Australia
Book Title: Electron Microscopy: Methods and Protocols
Series: Methods in Molecular Biology | Volume: 369 | Pub. Date: Feb-27-2007 | Page Range: 35-45 | DOI: 10.1007/978-1-59745-294-6_3
Subject: Biochemistry
Key Words: Plant tissue - ultrastructure - chemical fixation - dehydration - vacuum infiltration - resin embedding - cell wall - vacuole
Comments (Loading...) |
||
Loading... |





















