Cryo-electron tomography pipeline for plasma membranes.

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. © 2025. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.

Citation

Willy W Sun, Dennis J Michalak, Kem A Sochacki, Prasanthi Kunamaneni, Marco A Alfonzo-Méndez, Andreas M Arnold, Marie-Paule Strub, Jenny E Hinshaw, Justin W Taraska. Cryo-electron tomography pipeline for plasma membranes. Nature communications. 2025 Jan 20;16(1):855

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PMID: 39833141

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