biostudies-literatureFu XNCRR NIH HHSNIGMS NIH HHS1875-1882https://www.ebi.ac.uk/biostudies/studies/S-EPMC4255137biostudies-literatureUnknown22(12)Cryo-electron tomography (cryoET) has become a powerful tool for direct visualization of 3D structures of native biological specimens at molecular resolution, but its application is limited to thin specimens (<300 nm). Recently, vitreous sectioning and cryoFIB milling technologies were developed to physically reduce the specimen thickness; however, cryoET analysis of membrane protein complexes within native cell membranes remains a great challenge. Here, we use phage ΦX174 lysis gene E to rapidly produce native, intact, bacterial cell membranes for high resolution cryoET. We characterized E gene-induced cell lysis using FIB/SEM and cryoEM and showed that the bacteria cytoplasm was largely depleted through spot lesion, producing ghosts with the cell membranes intact. We further demonstratedStructure (London, England : 1993)Controlled bacterial lysis for electron tomography of native cell membranes.PMC4255137C06 RR017528-01GM085043C06 RR017528RR024424R01 GM085043R21 RR024424S10 RR029300Himes BAFu XRice WJNing JKe DZhang PfalseControlled bacterial lysis for electron tomography of native cell membranes.Cryo-electron tomography (cryoET) has become a powerful tool for direct visualization of 3D structures of native biological specimens at molecular resolution, but its application is limited to thin specimens (<300 nm). Recently, vitreous sectioning and cryoFIB milling technologies were developed to physically reduce the specimen thickness; however, cryoET analysis of membrane protein complexes within native cell membranes remains a great challenge. Here, we use phage ΦX174 lysis gene E to rapidly produce native, intact, bacterial cell membranes for high resolution cryoET. We characterized E gene-induced cell lysis using FIB/SEM and cryoEM and showed that the bacteria cytoplasm was largely depleted through spot lesion, producing ghosts with the cell membranes intact. We further demonstrated2014-01-01T00:00:00Z2014 Dec2025-06-27T03:05:31.661Z2025-06-27T03:05:31.661ZS-EPMC42551372545641310.1016/j.str.2014.09.017