chapter  five
9 Pages

Histological techniques

ByRobert N. Trigiano, Kathleen R. Malueg, Effin T. Graham

This chapter provides a "primer" of sorts for preparing specimens from plant tissue cultures for histological examination using elementary paraffin and scanning electron microscopy (SEM) techniques. It describes a procedure which works well with most tissues, including those from stock plants as well as those cultured in vitro. Moreover, it has several important advantages over more traditional paraffin protocols. First, it circumvents the use of toxic aldehydes and heavy metals as fixatives. Second, it avoids the use of specimen/slide adhesive agents, such as Haupt's, that require the use of formaldehyde and may produce staining artefacts. Third, it does not use toxic substances, such as xylene, for deparaffinizing sections. Fourth, sections are stained directly through the paraffin, greatly reducing the number of operational steps in process. The sections prepared for paraffin histology present two-dimensional interior view of a small area of much larger three-dimensional object. In contrast, scanning electron microscopy allows for three-dimensional topical or internal views of an entire specimen.