Scanning transmission electron microscopy through-focal tilt-series on biological specimens

•Three tilt-series schemes are compared: through-focus, combined and classic.•Acquisition protocol intrinsically contains three tilt-series schemes.•Reconstructions from the three tilt-series schemes can be compared pixel by pixel.•Through-focus reconstructions have highest contrast and amount of details.•Depth of field is generated by under- and over-focus images.