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

Abstract : 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 mi-croscope (FIB-SEM), offering high-resolution 3D imaging of large volumes andfields-of-view are becoming widely used in the life sciences. FIB-SEM has mostrecently been implemented on fully hydrated, cryo-immobilized, biological sam-ples. Correlative light and electron microscopy workflows combining fluores-cence microscopy (FM) with FIB-SEM imaging exist, whereas workflowscombining cryo-FM and cryo-FIB-SEM imaging are not yet commonly available.Here, we demonstrate that fluorescently labeled lipid droplets can serve asinsitufiducial markers for correlating cryo-FM and FIB-SEM datasets and that thisapproach can be used to target the acquisition of large FIB-SEM stacks spanningtens of microns under cryogenic conditions. We also show that cryo-FIB-SEM im-aging is particularly informative for questions related to organelle structure andinter-organellar contacts, nuclear organization, and mineral deposits in cells.
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http://hal.univ-nantes.fr/hal-03278159
Contributor : Perrine Paul-Gilloteaux <>
Submitted on : Monday, July 5, 2021 - 1:48:45 PM
Last modification on : Monday, July 12, 2021 - 12:03:33 PM

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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. iScience, Elsevier, 2021, 24 (7), pp.102714. ⟨10.1016/j.isci.2021.102714⟩. ⟨hal-03278159⟩

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