The sections (250 nm) were acquired using a Leica EM UC7 ultramicrotome (Leica) and collected on Formvar-coated copper slot grids, as previously described (Sun et al., 2021 (link)). After that, staining was performed with 2% uranyl acetate in 70% methanol and then with 0.4% lead citrate (17900; EMS). Commercial 15-nm gold particles (EMGC15; BBI Solutions) were added to both sides of the sections as fiducial markers. Dual tilt-axis series ranging from −60° to +60° were collected using an FEI Tecnai F20 electron microscope (Thermo Fisher Scientific), which was equipped with a Gatan US4000 (895) CCD camera and controlled with the FEI Xplore 3D TEM tomography software.
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Electron Tomography Sample Preparation
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Corresponding Organization : Tsinghua University
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Variable analysis
independent variables
- Acquisition method (Leica EM UC7 ultramicrotome)
- Staining procedure (2% uranyl acetate in 70% methanol, 0.4% lead citrate)
- Fiducial markers (15-nm gold particles)
- Dual tilt-axis series ranging from −60° to +60°
- Formvar-coated copper slot grids
- FEI Tecnai F20 electron microscope equipped with a Gatan US4000 (895) CCD camera and controlled with the FEI Xplore 3D TEM tomography software
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