Cryo-electron tomography of a cryo-FIB lift-out from adult mouse cortex

This dataset contains in situ cryo-electron tomography of vitreous adult mouse (C57BL/6J) cortical tissue, acquired to assess the quality of brain cryo-fixation achieved by a targeted molecular-biopsy workflow. A 200 um sagittal slice from a 144-day-old female mouse was biopsy-punched directly from the cortex, incubated for 30 min with a dextran/sucrose cryoprotectant, and vitrified by high-pressure freezing. Serial cryo-lift-out sections were excavated by xenon plasma focused-ion-beam (PFIB) milling, welded onto TEM support grids and thinned to ~150-300 nm lamellae with a final argon polish, revealing myelin, synaptic vesicles, mitochondria, membranes, ribosomes and microtubules together with the extracellular space preserved by vitrification. Tilt series were acquired on a Titan Krios G4 (300 kV) with a Selectris energy filter and Falcon 4i detector.