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Open AccessArticle
A Simple and User-Friendly Method for High-Quality Preparation of Pollen Grains for Scanning Electron Microscopy (SEM)
1
Center of Molecular Biotechnology, Russian State Agrarian University—Moscow Timiryazev Agricultural Academy, 49 Timiryazevskaya Str., Moscow 127550, Russia
2
All-Russian Research Institute of Agricultural Biotechnology, 42 Timiryazevskaya Str., Moscow 127550, Russia
3
Faculty of Fundamental Medicine, M. V. Lomonosov Moscow State University, Moscow 119991, Russia
4
Institute for Biomedical Problems, Russian Academy of Sciences, Moscow 123007, Russia
*
Author to whom correspondence should be addressed.
Plants 2024, 13(15), 2140; https://doi.org/10.3390/plants13152140 (registering DOI)
Submission received: 21 June 2024
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Revised: 28 July 2024
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Accepted: 31 July 2024
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Published: 1 August 2024
Abstract
Pollen is becoming an increasingly important subject for molecular researchers in genetic engineering, plant breeding, and environmental monitoring. To broaden the scope of these studies, it is essential to develop accessible methods for scientists who are not specialized in palynology. The article presents a simplified technical procedure for preparing pollen grains for scanning electron microscopy (SEM). The protocol is convenient for any molecular laboratory due to its small set of reagents, ease of execution, low cost, does not require special equipment, and takes only one hour to complete. The high penetrating ability of formaldehyde and the final delicate dehydration using hexamethyldisilazane (HMDS) instead of critical point drying allow for sufficient preservation of the architecture of the aperture, which is considered a gateway for the passage of biomolecules. The method was successfully applied to pollen grains of representatives of dicotyledons (beetroot, petunia, radish, tomato and tobacco) and monocotyledons (lily, onion, corn, rye and wheat). Species studied included insect-pollinated (entomophilous) and wind-pollinated (anemophilous) species. A comparative analysis of the sizes of fresh living pollen grains under a light microscope and those prepared for SEM showed some shrinkage. Quantitative analysis of the degree of pollen grain shrinkage showed that this process depends on the initial shape of dry pollen grains, and the number and structure of apertures. The results support the theoretical model of the folding/unfolding pathways of pollen grains.
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MDPI and ACS Style
Ermolaev, A.; Mardini, M.; Buravkov, S.; Kudryavtseva, N.; Khrustaleva, L.
A Simple and User-Friendly Method for High-Quality Preparation of Pollen Grains for Scanning Electron Microscopy (SEM). Plants 2024, 13, 2140.
https://doi.org/10.3390/plants13152140
AMA Style
Ermolaev A, Mardini M, Buravkov S, Kudryavtseva N, Khrustaleva L.
A Simple and User-Friendly Method for High-Quality Preparation of Pollen Grains for Scanning Electron Microscopy (SEM). Plants. 2024; 13(15):2140.
https://doi.org/10.3390/plants13152140
Chicago/Turabian Style
Ermolaev, Aleksey, Majd Mardini, Sergey Buravkov, Natalya Kudryavtseva, and Ludmila Khrustaleva.
2024. "A Simple and User-Friendly Method for High-Quality Preparation of Pollen Grains for Scanning Electron Microscopy (SEM)" Plants 13, no. 15: 2140.
https://doi.org/10.3390/plants13152140
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