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 (Kaur and Kariyat, 2020a, 2020b; Watts and Kariyat, 2021), 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 Scanning Electron Microscope (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.
Sakshi Watts, Ishveen Kaur, Sukhman Singh, Bianca Jimenez, Jesus Chavana, Rupesh Kariyat, Desktop scanning electron microscopy in plant-insect interactions research: A fast and effective way to capture electron micrographs with minimal sample preparation, Biology Methods and Protocols, 2021;, bpab020, https://doi.org/10.1093/biomethods/bpab020
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Biology Methods and Protocols