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

Abstract

Abstract 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.

Plant phenotyping relevance

植物表面形態（葉毛・ワックス等）の取得を目的に、デスクトップSEMの簡便な試料調製・撮像法を開発し、実例で有効性を検証しているため。

Code and data availability

The article is a methods paper on desktop SEM imaging of plant–insect samples. No public phenotype dataset, image repository, analysis code, or model deposit is mentioned; the data availability statement only states the data are in the article itself, and no authors' public URL for assets is provided.

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