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Desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation

The ability to visualize cell and tissue morphology at a high magnification using scanning electron microscopy (SEM) has revolutionized plant sciences research. In plant–insect interactions studies, SEM-based imaging has been of immense assistance to understand plant surface morphology including tri...

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Detalles Bibliográficos
Autores principales: Watts, Sakshi, Kaur, Ishveen, Singh, Sukhman, Jimenez, Bianca, Chavana, Jesus, Kariyat, Rupesh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8754489/
https://www.ncbi.nlm.nih.gov/pubmed/35036571
http://dx.doi.org/10.1093/biomethods/bpab020
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author Watts, Sakshi
Kaur, Ishveen
Singh, Sukhman
Jimenez, Bianca
Chavana, Jesus
Kariyat, Rupesh
author_facet Watts, Sakshi
Kaur, Ishveen
Singh, Sukhman
Jimenez, Bianca
Chavana, Jesus
Kariyat, Rupesh
author_sort Watts, Sakshi
collection PubMed
description The ability to visualize cell and tissue morphology at a high magnification using scanning electron microscopy (SEM) has revolutionized plant sciences research. In plant–insect interactions studies, SEM-based imaging has been of immense assistance to understand plant surface morphology including trichomes [plant hairs; physical defense structures against herbivores], spines, waxes, and insect morphological characteristics such as mouth parts, antennae, and legs, that they interact with. While SEM provides finer details of samples, and the imaging process is simpler now with advanced image acquisition and processing, sample preparation methodology has lagged. The need to undergo elaborate sample preparation with cryogenic freezing, multiple alcohol washes, and sputter coating makes SEM imaging expensive, time consuming, and warrants skilled professionals, making it inaccessible to majority of scientists. Here, using a desktop version of SEM (SNE- 4500 Plus Tabletop), we show that the “plug and play” method can efficiently produce SEM images with sufficient details for most morphological studies in plant–insect interactions. We used leaf trichomes of Solanum genus as our primary model, and oviposition by tobacco hornworm (Manduca sexta; Lepidoptera: Sphingidae) and fall armyworm (Spodoptera frugiperda; Lepidoptera: Noctuidae), and leaf surface wax imaging as additional examples to show the effectiveness of this instrument and present a detailed methodology to produce the best results with this instrument. While traditional sample preparation can still produce better resolved images with less distortion, we show that even at a higher magnification, the desktop SEM can deliver quality images. Overall, this study provides detailed methodology with a simpler “no sample preparation” technique for scanning fresh biological samples without the use of any additional chemicals and machinery.
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spelling pubmed-87544892022-01-13 Desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation Watts, Sakshi Kaur, Ishveen Singh, Sukhman Jimenez, Bianca Chavana, Jesus Kariyat, Rupesh Biol Methods Protoc Methods Article The ability to visualize cell and tissue morphology at a high magnification using scanning electron microscopy (SEM) has revolutionized plant sciences research. In plant–insect interactions studies, SEM-based imaging has been of immense assistance to understand plant surface morphology including trichomes [plant hairs; physical defense structures against herbivores], spines, waxes, and insect morphological characteristics such as mouth parts, antennae, and legs, that they interact with. While SEM provides finer details of samples, and the imaging process is simpler now with advanced image acquisition and processing, sample preparation methodology has lagged. The need to undergo elaborate sample preparation with cryogenic freezing, multiple alcohol washes, and sputter coating makes SEM imaging expensive, time consuming, and warrants skilled professionals, making it inaccessible to majority of scientists. Here, using a desktop version of SEM (SNE- 4500 Plus Tabletop), we show that the “plug and play” method can efficiently produce SEM images with sufficient details for most morphological studies in plant–insect interactions. We used leaf trichomes of Solanum genus as our primary model, and oviposition by tobacco hornworm (Manduca sexta; Lepidoptera: Sphingidae) and fall armyworm (Spodoptera frugiperda; Lepidoptera: Noctuidae), and leaf surface wax imaging as additional examples to show the effectiveness of this instrument and present a detailed methodology to produce the best results with this instrument. While traditional sample preparation can still produce better resolved images with less distortion, we show that even at a higher magnification, the desktop SEM can deliver quality images. Overall, this study provides detailed methodology with a simpler “no sample preparation” technique for scanning fresh biological samples without the use of any additional chemicals and machinery. Oxford University Press 2021-10-13 /pmc/articles/PMC8754489/ /pubmed/35036571 http://dx.doi.org/10.1093/biomethods/bpab020 Text en © The Author(s) 2021. Published by Oxford University Press. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Methods Article
Watts, Sakshi
Kaur, Ishveen
Singh, Sukhman
Jimenez, Bianca
Chavana, Jesus
Kariyat, Rupesh
Desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation
title Desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation
title_full Desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation
title_fullStr Desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation
title_full_unstemmed Desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation
title_short Desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation
title_sort desktop scanning electron microscopy in plant–insect interactions research: a fast and effective way to capture electron micrographs with minimal sample preparation
topic Methods Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8754489/
https://www.ncbi.nlm.nih.gov/pubmed/35036571
http://dx.doi.org/10.1093/biomethods/bpab020
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