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. Author manuscript; available in PMC: 2021 Aug 8.
Published in final edited form as: J Struct Biol. 2018 Feb 1;202(3):216–228. doi: 10.1016/j.jsb.2018.01.012

Table 1.

Comparison of sample preparation, data acquisition, and data processing between STEM tomography and SBF-SEM

Parameter╲Technique Serial block face SEM BF-STEM tomography
Cell / tissue volume ~ 103 – 106 μm3 ~ 101 – 102 μm3
Specimen shape solid block thick sections cut to thickness of 1 – 2 μm
Acquisition mode image stack dual-axis tilt series (~ ±70°) with ~2° angular increment
Specimen preparation conventional fixation or freeze substitution; heavy staining with Os, U, Pb; plastic embedding conventional fixation or freeze-substitution; light staining with Os, U; plastic embedding
Incident electron energy 1.0 – 1.5 keV 200 – 300 keV
Minimum voxel size 5 nm in x, y 25 nm in z 3 – 5 nm approximately isotropic
Acquisition time ~ 103 −105 s ~ 102 −103 s
Processing time for reconstruction — ~ 103 – 104 s
Reconstruction method — weighted back-projection SIRT, compressed sensing
Processing time for segmentation ~ 105 −106 s ~ 104 −105 s
Electron fluence < 20 electrons / nm2 < 104 electrons / nm2 after initial pre-irradiation
Effect of electron fluence shrinkage of block during acquisition of image stack; irregular cutting shrinkage normal to section plane prior to acquisition of tilt series; shortened voxels along z -axis
Artifacts image processing electrical charging of block missing wedge or missing pyramid; reconstruction errors