Steedman's Wax Sectioning for Observation of the Microtubule Cytoskeleton in Plant Cells

As one of the major components of the cytoskeleton, the microtubule cytoskeleton plays an important role in plant development. The present paper gives a brief introduction on the techniques widely applied in this field and their limitations, and recommends a method, a low melting polyester wax (steedman's wax) as an embedding medium for the investigation of the microtubule cytoskeleton with immuno-cytochemistry. Its low melt point (35-37℃) makes it favorable for keeping the antigenicity. Chlorophyll is extracted in the process of infiltration so that the interference of intense autofluorescence from chloroplasts decreases. Steedman's wax has similar characters with paraffin, being able to cut into ribbon-sections of different thickness, which is beneficial both to antibody penetration because of the breaking of cell wall and to the observation of successive focal planes (in ribbon-sections) with confocal laser scanning microscopy. General procedures of Steedman's wax sectioning and some practical improvements were also discussed in the present paper.