Nano-injection method for micro-insects without sedation using the western flower thrips, frankliniella occidentalis
2022
Raffin, Catherine Lee | Price, Briana Elizabeth | Yun, Seung Hwan | Choi, Man Yeon
Micro-injection techniques are invaluable in entomological research as they allow for the direct delivery of biological compounds into the hemocoel of specimens at controlled volumes. Nearly all micro-injection methods (> 4 mm in size) utilize forcibly immobilized insects that could affect specimen fitness and physiology. This alteration can cause inconclusive interpretation of phenotypic reactions post-injection. In this study, we detail a novel, improved nano-volume injection technique for micro-insects (< 3 mm) using the western flower thrips, Frankliniella occidentallis as a model. By constructing and using a modified vacuum stand, injections were performed on live thrips, without sedation and physical injury. A simple survivorship assay in which the thrips were injected with 10 nl water into the thorax or abdomen demonstrated that thoracic injections yield similar survival rates to control thrips that were not injected, while abdominal injections severely limited survivorship. Here we present the integrative method that customizes the vacuum stand, nano-injection tools, injecting volume, and other specifications for live micro-insects.
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