Sample preparation including dehydration and drying of samples is the most intricate part of scanning electron microscopy. Most current sample preparation protocols use critical-point drying with liquid carbon dioxide. Very few studies have reported samples that were dried using chemical reagents. In this study, we used hexamethyldisilazane, a chemical drying reagent, to prepare plant samples. As glandular trichomes are among the most fragile and sensitive surface structures found on plants, we used Millingtonia hortensis leaf samples as our study materials because they contain abundant glandular trichomes. The results obtained using this new method are identical to those produced via critical-point drying.

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Sample preparation including the dehydration and drying of the samples is the most intricate part in the field of scanning electron microscopy. Till to date, most of the sample preparation protocols use critical point drying with the help of liquid carbon dioxide. Very few cases have been reported where samples were dried using some chemical reagents. In this work, we have explored the possibility of using hexamethyldisilazane, a chemical drying reagent, for the preparation of plant samples. As glandular trichomes are one of the most fragile and sensitive surface structures found on plants, we chose to use Millingtonia hortensis leaf samples as our study materials that contain abundantly glandular trichome. The results obtained by this new method are identical with the results obtained from the critical point drying method, in every aspect.

Polyethyleneglycol (PEG) is considered one of the most effective substitutions for paraffin in plant histochemistry as an embedding medium. A rapid and straightforward method of PEG embedding has been developed that resulted in a significant reduction of infiltration time than the traditional method of PEG embedding. The material used for PEG embedding was leaves of Cinnamomum tamala, a member of Lauraceae. Samples were put successively in aqueous solutions of PEG 6000 with increasing concentration for infiltration. A microwave oven was used as a mode of heating medium. The infiltration was completed within 2 h. After the completion of infiltration, the samples were embedded in PEG and solidified. Compared with the existing methods available for PEG infiltration and embedding, this microwave mediated PEG embedding method saves significant time; this also saves the tissue from long-term heat-induced damage. Retention of intracellular metabolites, which was not possible in earlier PEG embedded methods, has also demonstrated in the tissue.

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