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TitleLift-out cryo-FIBSEM and cryo-ET reveal the ultrastructural landscape of extracellular matrix.
Journal, issue, pagesJ Cell Biol, Vol. 223, Issue 6, Year 2024
Publish dateJun 3, 2024
AuthorsBettina Zens / Florian Fäßler / Jesse M Hansen / Robert Hauschild / Julia Datler / Victor-Valentin Hodirnau / Vanessa Zheden / Jonna Alanko / Michael Sixt / Florian K M Schur /
PubMed AbstractThe 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 ...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.
External linksJ Cell Biol / PubMed:38506714 / PubMed Central
MethodsEM (tomography)
Structure data

EMDB-18490: Cryo-electron tomogram of lift-out lamella from cell-derived extracellular matrix (example 1)
Method: EM (tomography)

EMDB-18491: Cryo-electron tomogram of lift-out lamella from cell-derived extracellular matrix (example 2)
Method: EM (tomography)

EMDB-18492: Cryo-electron tomogram of lift-out lamella from cell-derived extracellular matrix (example 3)
Method: EM (tomography)

EMDB-18493: Cryo-electron tomogram of lift-out lamella from cell-derived extracellular matrix (example 4)
Method: EM (tomography)

EMDB-18494: Cryo-electron tomogram of lift-out lamella from cell-derived extracellular matrix (example 5)
Method: EM (tomography)

Source
  • Homo sapiens (human)

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