Extracellular filaments revealed by affinity capture cryogenic-electron tomography

Leeya Engel (), Magda Zaoralová, Momei Zhou, Alexander R. Dunn and Stefan L. Oliver ()
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Leeya Engel: Stanford University
Magda Zaoralová: Stanford University
Momei Zhou: Stanford University School of Medicine
Alexander R. Dunn: Stanford University
Stefan L. Oliver: SLAC National Accelerator Laboratory

Nature Communications, 2025, vol. 16, issue 1, 1-11

Abstract: Abstract 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.

Date: 2025
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DOI: 10.1038/s41467-025-64795-z

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