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Agenda


Séminaire invité IBS

Cryo-correlative light and electron tomography : a 3d glimpse into intact cells and tissues at molecular resolution

​Vendredi 07 février 2025 à 11:00, Salle de séminaire IBS, 71 avenue des Martyrs, Grenoble

Publié le 7 février 2025
Dr Anna Sartori-Rupp
Nanoimagerie, Institut Pasteur, Paris
In cellulo cryo-correlative light and electron microscopy (Cryo-CLEM) methods are powerful and complex method that widen our understanding of cellular processes at molecular resolution by linking functional and structural information on close to native samples. Specific structures of interest can be labelled fluorescently and then localised and imaged at molecular resolution in 3D within their unperturbed cellular environment by cryo-electron tomography. To image a cellular sample in an electron microscope, it must be milled to produce thin cryo-lamellae ( 200 nm), acting as electron transparent windows through the cell and nevertheless representing only a small portion of the overall cell volume. In this presentation I will illustrate the principle of cryo-CLEM methods and the advanced pipelines that we developed at Institut Pasteur with focus on host-pathogen interaction studies. These novel pipelines can be applied more generally to the structural study of biological process within a cell that can be targeted by fluorescent labelling. I will discuss the challenges of these methods related to the key step of fluorescent imaging in cryogenic conditions and of the fine registration of 3D cryo-FLM volumes with EM data. Finally, I will conclude by presenting the current available solutions and perspectives to improve the resolution of cryo-fluorescence microscopy and the targeting of labelled structures with sub-micron precision to address questions at the single cell level and beyond.​

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