Cryo-electron tomography pipeline for plasma membranes.

Sun, Willy W; Michalak, Dennis J; Sochacki, Kem A; Kunamaneni, Prasanthi; Alfonzo-Méndez, Marco A; Arnold, Andreas M; Strub, Marie-Paule; Hinshaw, Jenny E et al. · Nat Commun · 2025

basic_science · Level V

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Abstract

Cryo-electron tomography (cryoET) provides sub-nanometer protein structure within the dense cellular environment. Existing sample preparation methods are insufficient at accessing the plasma membrane and its associated proteins. Here, we present a correlative cryo-electron tomography pipeline optimally suited to image large ultra-thin areas of isolated basal and apical plasma membranes. The pipeline allows for angstrom-scale structure determination with subtomogram averaging and employs a genetically encodable rapid chemically-induced electron microscopy visible tag for marking specific proteins within the complex cellular environment. The pipeline provides efficient, distributable, low-cost sample preparation and enables targeted structural studies of identified proteins at the plasma membrane of mammalian cells.

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