Cryo-electron tomography pipeline for plasma membranes

Cryo-electron tomography pipeline for plasma membranes

Blog Article

Abstract Cryo-electron tomography (cryoET) provides sub-nanometer protein structure within the Floor Sweepers 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 Wind Chimes 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.