Know more

The microscopist's dilemma is that he'd like to have it all, i.e. observe large volumes, at high spatial and temporal resolution, and as close as possible to the structure's native state! In order to answer these wishes, it is necessary to consider the most important questions the microscopist wants to answer for a given experiment, such as "do I need a large volume or high resolution? do I need temporal or structural information? etc.".

Correlative Light and Electron Microscopy (CLEM) combines the advantages of optical and electron microscopy, to locate and visualize cells or macromolecular structures.

Pulmonary tissue is complex and very difficult to prepare for transmission electron microscopy (TEM), making the study of its constituents and the localization of proteins all the more complicated. Pulmonary hypertension is a serious, lethal disease involving small pulmonary arteries (<500 µm). The expression and localization of proteins involved in this pathology would help to understand the biological mechanisms; phospho-Vimentin (P-Vim) is one such protein of interest, but difficult to localize using conventional immunofluorescence techniques. 

The Cellular Ultrastructure Unit (PUC) of the MIMA2 platform has specifically developed a method for animal tissues, enabling preservation of these tissues and of arterial cell morphology, as well as signal preservation. This methodology enabled P-Vim to be localized to pulmonary endothelial and sub-endothelial cells, which are not identifiable using light microscopy. Fluorescence microscopy and TEM images are then correlated to pinpoint areas of interest.

Contact :

• Christine Péchoux 

Reference :
Christine Péchoux, Fabrice Antigny, Frédéric Perros,
A correlated light and electron microscopy approach to study the subcellular localization of phosphorylated vimentin in human lung tissue. Methods in Cell Biology, Academic Press, 2024,
ISSN 0091-679X,
https://doi.org/10.1016/bs.mcb.2024.02.034.
(https://www.sciencedirect.com/science/article/pii/S0091679X24000657)