Minimal resin embedding of multicellular specimens for targeted FIB-SEM imaging

Methods in Cell Biology
Nicole L SchieberAnna M Steyer

Abstract

Correlative light and electron microscopy (CLEM) is a powerful tool to perform ultrastructural analysis of targeted tissues or cells. The large field of view of the light microscope (LM) enables quick and efficient surveys of the whole specimen. It is also compatible with live imaging, giving access to functional assays. CLEM protocols take advantage of the features to efficiently retrace the position of targeted sites when switching from one modality to the other. They more often rely on anatomical cues that are visible both by light and electron microscopy. We present here a simple workflow where multicellular specimens are embedded in minimal amounts of resin, exposing their surface topology that can be imaged by scanning electron microscopy (SEM). LM and SEM both benefit from a large field of view that can cover whole model organisms. As a result, targeting specific anatomic locations by focused ion beam-SEM (FIB-SEM) tomography becomes straightforward. We illustrate this application on three different model organisms, used in our laboratory: the zebrafish embryo Danio rerio, the marine worm Platynereis dumerilii, and the dauer larva of the nematode Caenorhabditis elegans. Here we focus on the experimental steps to reduce t...Continue Reading

Citations

Sep 25, 2018·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Manja Luckner, Gerhard Wanner
May 24, 2018·Histochemistry and Cell Biology·Manja Luckner, Gerhard Wanner
Dec 9, 2020·Communications Biology·Jean-Claude TinguelyBalpreet Singh Ahluwalia
Aug 12, 2021·Cell·Hernando M VergaraDetlev Arendt

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