In-situ fiducial markers for 3D correlative cryo- fluorescence and FIB-SEM imaging - Archive ouverte HAL Accéder directement au contenu
Pré-Publication, Document De Travail Année : 2021

In-situ fiducial markers for 3D correlative cryo- fluorescence and FIB-SEM imaging

Résumé

Imaging of cells and tissues has improved significantly over the last decade. Dual-beam instruments with a focused ion beam mounted on a scanning electron microscope (FIB-SEM), which offer high-resolution 3D imaging of large volumes and fields-of-view are becoming widely used in the life sciences. FIB-SEM has most recently been implemented on fully hydrated, cryo-immobilized, biological samples. However, correlative light and electron microscopy (CLEM) workflows combining cryo- fluorescence microscopy (cryo-FM) and FIB-SEM are not yet commonly available. Here, we demonstrate that fluorescently labeled lipid droplets can serve as in-situ fiducial markers for correlating cryo- FM and FIB-SEM datasets, and that this approach can be used to target the acquisition of large FIB-SEM stacks spanning tens of microns under cryogenic conditions. We also show that cryo-FIB-SEM imaging is particularly informative for questions related to organelle structure and inter-organellar contacts, nuclear organization and mineral deposits in cells.

Dates et versions

hal-03171819 , version 1 (17-03-2021)

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Nadav Scher, Katya Rechav, Perrine Paul-Gilloteaux, Ori Avinoam. In-situ fiducial markers for 3D correlative cryo- fluorescence and FIB-SEM imaging. 2021. ⟨hal-03171819⟩
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