Biology Reference
In-Depth Information
Chapter 1
Essential Methods of Plant Sample Preparation
for Light Microscopy
Aleš Soukup and Edita Tylová
Abstract
There are various preparatory techniques for light microscopy permitting access to the inner structure of
plant body and its development. Minute objects might be processed as whole-mount preparations, while
voluminous ones should be separated into smaller pieces. Hereby we summarize some of the “classical”
techniques to cut more voluminous objects into slices and access their inner structure either for simple
anatomical analysis or for further processing (e.g., histochemistry, immunohistochemistry, in situ hybrid-
ization, enzyme histochemistry).
Key words
Paraffi n, Sections, Freehand sectioning, Fixation, Whole-mount, Serial sections,
Cryotome, Hand microtome
1
Introduction
There are various ways of preparation of plant objects to be
investigated with light microscopy. Correct selection of appropri-
ate technique largely depends on equipment available, but nature,
optical character, complexity, and size of the object and purpose of
the preparation take major part. Hereby we present a set of simple
techniques which might provide vast, however not exhaustive,
information on structural and cytological features of cells, tissues,
and organs. Tissues, organs, or explants, which are not voluminous
and optically dense, might be processed as a cleared whole-mount
preparations. Such a way of preparation became very popular with
advent of confocal microscopy and Arabidopsis as a model plant.
However, there are many objects where cuttings or macerations
are necessary to gain adequate information on internal structure.
Available sectioning techniques allow preparation of sections with
variable thickness according to intended application. Tissue preser-
vation (fi xation) and embedding into supporting matrix are com-
mon initial steps involved in most of sectioning methods
