A1 Refereed original research article in a scientific journal
Lift-out cryo-FIBSEM and cryo-ET reveal the ultrastructural landscape of extracellular matrix
Authors: Zens, Bettina; Fäßler, Florian; Hansen, Jesse M.; Hauschild, Robert; Datler, Julia; Hodirnau, Victor-Valentin; Zheden, Vanessa; Alanko, Jonna; Sixt, Michael; Schur, Florian K.M.
Publisher: Rockefeller University Press
Publication year: 2024
Journal: Journal of Cell Biology
Journal name in source: Journal of Cell Biology
Volume: 223
ISSN: 0021-9525
eISSN: 1540-8140
DOI: https://doi.org/10.1083/jcb.202309125
Web address : https://doi.org/10.1083/jcb.202309125
The extracellular matrix (ECM) serves as a scaffold for cells and plays an essential role in regulating numerous cellular processes, including cell migration and proliferation. Due to limitations in specimen preparation for conventional room-temperature electron microscopy, we lack structural knowledge on how ECM components are secreted, remodeled, and interact with surrounding cells. We have developed a 3D-ECM platform compatible with sample thinning by cryo-focused ion beam milling, the lift-out extraction procedure, and cryo-electron tomography. Our workflow implements cell-derived matrices (CDMs) grown on EM grids, resulting in a versatile tool closely mimicking ECM environments. This allows us to visualize ECM for the first time in its hydrated, native context. Our data reveal an intricate network of extracellular fibers, their positioning relative to matrix-secreting cells, and previously unresolved structural entities. Our workflow and results add to the structural atlas of the ECM, providing novel insights into its secretion and assembly.