Browse > Article
http://dx.doi.org/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 (Natural Product Biotechnology Group, Agricultural and Food Engineering Department, Indian Institute of Technology Kharagpur)
Sulagna Saha (Natural Product Biotechnology Group, Agricultural and Food Engineering Department, Indian Institute of Technology Kharagpur)
Olga Kostina (Laboratory of Plant Anatomy and Morphology, Komarov Botanical Institute, Russian Academy of Sciences)
Lyudmila Muravnik (Laboratory of Plant Anatomy and Morphology, Komarov Botanical Institute, Russian Academy of Sciences)
Adinpunya Mitra (Natural Product Biotechnology Group, Agricultural and Food Engineering Department, Indian Institute of Technology Kharagpur)
Publication Information
Applied Microscopy / v.50, no., 2020 , pp. 15.1-15.6 More about this Journal
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
Scanning electron microscope; Critical-point drying; Chemical drying; Hexamethyldisilazane; Millingtonia hortensis; Glandular trichomes;
Citations & Related Records
연도 인용수 순위
  • Reference
1 J.L. Nation, A new method using hexamethyldisilazane for preparation of soft insect tissues for scanning electron microscopy. Stain. Technol. 58, 347-351 (1983)
2 W. Chissoe, E.L. Vezey, J.J. Skvarla, Hexamethyldisilazane as a drying agent for pollen scanning electron microscopy. Biotech. Histochem. 69, 192-198 (1994)
3 W.T. Gunning, R.E. Crang, The usefulness of glutaraldehyde-carbohydrazide copolymerization in biological specimen stabilization for scanning electron microscopy. J. Electron Microsc. Tech. 1, 131-140 (1984)
4 S.J. Livingston, T.D. Quilichini, J.K. Booth, D.C.J. Wong, K.H. Rensing, J. Laflamme- Yonkman, S.D. Castellarin, J. Buhrmann, J.E. Page, A.L. Samuels, Cannabis glandular trichomes alter morphology and metabolite content during flower maturation. Plant J. 101, 37-56 (2020)
5 G.A. Meek, Practical electron microscopy for biologists, 2nd edn. (Wiley, London, 1976), pp. 387-423
6 L.E. Muravnik, O.V. Kostina, A.L. Shavarda, Glandular trichomes of Tussilago farfara (Senecioneae, Asteraceae). Planta 244, 737-752 (2016)
7 P. Schols, E. Koen, C. D'Hondt, V. Merckx, E. Smets, S. Huysmans, A new enzymebased method for the treatment of fragile pollen grains collected from herbarium materials. Taxon 56, 777-782 (2004)
8 M.R. Zuzarte, A.M. Dinisa, C. Cavaleiro, L.R. Salgueiro, J.M. Canhoto, Trichomes, essential oils and in vitro propagation of Lavandula pedunculata (Lamiaceae). Ind. Crop. Prod. 32, 580-587 (2010)
9 S.A. Nierzwicki-Baur, in Ultrastructure techniques for microorganisms, ed. by H. C. Aldrich, W. J. Todd. High voltage electron microscopy (Plenum Press, New York, 1986), pp. 297-332
10 R.J. Adams, J.K. Morton, An improved technique for examining pollen under the scanning electron microscope. Pollen Spores 14, 203-212 (1972)
11 F. Braet, D.E. Zanger, E. Wisse, Drying cells for SEM, AFM and TEM by hexamethyldisilazane: a study on hepatic endothelial cells. J. Microsc. 186, 84-87 (1997)
12 J.C. Araujo, F.C. Teran, R.A. Oliveira, E.A.A. Nour, M.A.P. Montenegro, J.R. Campos, R.F. Vazoller, Comparison of hexamethyldisilazane and critical-point drying treatments for SEM analysis of anaerobic biofilms and granular sludge. J. Electron Microsc. 52, 429-433 (2003)
13 K. Bomblies, V. Shukla, C. Graham, Scanning electron microscopy (SEM) of plant tissues. CSH Protocols (2008). https://doi.org/10.1101/pdb.prot4933   DOI
14 A. Boyde, in Electron microscopy, ed. by P. Brederoo, W. de Priester. Review of basic preparation techniques for biological scanning electron microscopy (Electron Microscopy Foundation, Leiden, 1980), pp. 768-777
15 D.F. Bray, J. Bagu, P. Koegler, Comparison of hexamethyldisilazane (HMDS), Peldri II, and critical-point drying methods for scanning electron microscopy of biological specimens. Microsc. Res. Tech. 26, 489-495 (1993)
16 A.K. Pathan, J. Bond, R.E. Gaskin, Sample preparation for SEM of plant surfaces. Mater. Today 12, 32-43 (2010)
17 S. Shively, W.R. Miller, The use of HMDS (hexamethyldisilazane) to replace criticalpoint drying (CPD) in the preparation of tardigrades for SEM (scanning electron microscope) imaging. Trans. Kans. Acad. Sci. 112, 198-200 (2009)
18 B. Zimmer, I.I. Peldri, A quick and easy alternative to critical-point drying for scanning electron microscopy. Am. Fern J. 79, 146-150 (1989)