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. 2020 Jul 17;50(1):15.
doi: 10.1186/s42649-020-00035-6.

Replacing critical point drying with a low-cost chemical drying provides comparable surface image quality of glandular trichomes from leaves of Millingtonia hortensis L. f. in scanning electron micrograph

Raktim Bhattacharya  1 , Sulagna Saha  1 , Olga Kostina  2 , Lyudmila Muravnik  2 , Adinpunya Mitra  3
Affiliations

Replacing critical point drying with a low-cost chemical drying provides comparable surface image quality of glandular trichomes from leaves of Millingtonia hortensis L. f. in scanning electron micrograph

Raktim Bhattacharya et al. Appl Microsc. .

Abstract

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.

Keywords: Chemical drying; Critical-point drying; Glandular trichomes; Hexamethyldisilazane; Millingtonia hortensis; Scanning electron microscope.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Scanning electron micrograph of the M. hortensis leaf surface after critical-point drying (a) without OsO4 and (b) with OsO4. Imaged at 7.00 kV with a magnification of X600. Scale bar represents 20 μm
Fig. 2
Fig. 2
Scanning electron micrograph of the M. hortensis leaf surface after HMDS-based drying (a) without OsO4 and (b) with OsO4. Imaged at 20.00 kV with a magnification of X500. Scale bar represents 30 μm
Fig. 3
Fig. 3
Scanning electron micrograph of the M. hortensis leaf surface after air drying (a) without OsO4, and (b) with OsO4. Imaged at 20.00 kV with a magnification of X500. Scale bar represents 20 μm
Fig. 4
Fig. 4
Scanning electron micrograph of the M. hortensis leaf surface after CPD (a and b), HMDS (c and d), and air drying (e and f). All the samples were treated with OsO4 as a secondary fixative while maintaining all of the sample preparation parameters, except for the final drying process. Scale bars represent 10 μm in a, b, and c and 3 μm in d, e, and f
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