Electron Microscopy of Plant Cells by C. Hawes (Eds.)

By C. Hawes (Eds.)

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The stain is made by dissolving 1 g of toluidine blue and 1 g of sodium borate (borax) in 100 ml of water (or in a 2 % solution of sodium carbonate) and filtering. The solution is stable. For staining, one large drop of the solution is placed on the sections stuck on a glass slide. Staining is progressive and directly perceptible. Staining time is shortened if the slide is warmed on a hot plate (5-15 min at 40-50°C, depending on the thickness of the section and the nature of the tissue). The stain is drained off and the slide rinsed with water (two successive immersions of 1-2 min in distilled water or rinsing with slow running tap water).

General view. Stony pericarp of hazel nut. ; pf, pit field; lu, lumen ( x 3000). C. Roland and B. Vian The technique generally does not differentiate between compounds of similar structure or isomers of the same compound. Nor does it provide information on monomer sequences, the distinction of which has until now been outside the scope of available LM and EM cytochemical techniques. Affinity detection has opened new avenues for cytochemistry. g. monomers, sequences, basic motifs). Affinity methods use three main types of probes: lectins, enzymes and antibodies.

The purification of numerous lectins has provided versatile new tools for topochemistry. g. molluscs and fishes). The key reference in the field is the Concanavalin A (Con A) purified from seeds of Canavalia ensiformis which recognizes terminal α-glucopyranosyl and amannopyranosyl residues (Goldstein et ah, 1978). It has been used, for example, to detect the glycocalyx coating the outer face of animal and plant plasma membranes. In recent years, the list of available lectins has rapidly increased giving a large spectrum of specific probes for sugars (Roth, 1983b; Benhamou, 1989).

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