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TitleExtracellular filaments revealed by affinity capture cryogenic-electron tomography.
Journal, issue, pagesNat Commun, Vol. 16, Issue 1, Page 9802, Year 2025
Publish dateNov 6, 2025
AuthorsLeeya Engel / Magda Zaoralová / Momei Zhou / Alexander R Dunn / Stefan L Oliver /
PubMed AbstractCryogenic-electron tomography (cryo-ET) has provided an unprecedented glimpse into the nanoscale architecture of cells by combining cryogenic preservation of biological structures with electron ...Cryogenic-electron tomography (cryo-ET) has provided an unprecedented glimpse into the nanoscale architecture of cells by combining cryogenic preservation of biological structures with electron tomography. Micropatterning of extracellular matrix proteins is increasingly used as a method to prepare adherent cell types for cryo-ET as it promotes optimal positioning of cells and subcellular regions of interest for vitrification, cryo-focused ion beam (cryo-FIB) milling, and data acquisition. Here we demonstrate a micropatterning workflow for capturing minimally adherent cell types, human T cells and Jurkat cells, for cryo-FIB and cryo-ET. Our affinity capture system facilitated the nanoscale imaging of Jurkat cells, revealing extracellular filamentous structures. It improved workflow efficiency by consistently producing grids with a sufficient number of well-positioned cells for an entire cryo-FIB session. Affinity capture can be extended to facilitate high-resolution imaging of other adherent and non-adherent cell types with cryo-ET.
External linksNat Commun / PubMed:41198659 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution10.8 Å
Structure data

EMDB-43978: Subtomogram average of extracellular intermediate filaments from Jurkat cells
Method: EM (subtomogram averaging) / Resolution: 10.8 Å

Source
  • Homo sapiens (human)

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