A1 Refereed original research article in a scientific journal

Lift-out cryo-FIBSEM and cryo-ET reveal the ultrastructural landscape of extracellular matrix




AuthorsZens, Bettina; Fäßler, Florian; Hansen, Jesse M.; Hauschild, Robert; Datler, Julia; Hodirnau, Victor-Valentin; Zheden, Vanessa; Alanko, Jonna; Sixt, Michael; Schur, Florian K.M.

PublisherRockefeller University Press

Publication year2024

JournalJournal of Cell Biology

Journal name in sourceJournal of Cell Biology

Volume223

ISSN0021-9525

eISSN1540-8140

DOIhttps://doi.org/10.1083/jcb.202309125

Web address https://doi.org/10.1083/jcb.202309125


Abstract

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.



Last updated on 2025-27-01 at 19:07