A069 - In Resin Fluorescence (IRF) Correlative Light and Electron Microscopy (CLEM) for plant tissues

Clement Chambaud (clement.chambaud@u-bordeaux.fr)
Amandine Leroy (amandine.leroy@inrae.fr)

Correlative Light and Electron Microscopy (CLEM) consists in imaging the very same cell or tissue area using first light followed by electron microscopy. CLEM approaches permit to identify the region of interest labelled by fluorescence and to access to the ultrastructural details of this region. Here, we will focus on one of them called In Resin Fluorescence (IRF) that we adapted for plant tissues. After sample preparation and sectioning, a tricky steps consist to catch enough fluorescent signal before ultrastructure observations. After introducing main principles of sample preparation for IRF, we propose to show tips and tricks allowing a correct light microscopy acquisitions with the help of Airyscan to get the most out of the fluorescence microscopy. We will finish by showing how to overlay light and electron acquisitions thanks to the free software ICY and the plugin EcCLEM.