A quantitative four-dimensional scanning transmission electron microscopy (4D-STEM) imaging technique (q4STEM) for local thickness estimation across amorphous specimen such as obtained by focused ion beam (FIB)-milling of lamellae for (cryo-)TEM analysis is presented. This study is based on measuring spatially resolved diffraction patterns to obtain the angular distribution of electron scattering, or the ratio of integrated virtual dark and bright field STEM signals, and their quantitative evaluation using Monte Carlo simulations. The method is independent of signal intensity calibrations and only requires knowledge of the detector geometry, which is invariant for a given instrument. This study demonstrates that the method yields robust thickness estimates for sub-micrometer amorphous specimen using both direct detection and light conversion 2D-STEM detectors in a coincident FIB-SEM and a conventional SEM. Due to its facile implementation and minimal dose reauirements, it is anticipated that this method will find applications for in situ thickness monitoring during lamella fabrication of beam-sensitive materials.

Department of Bionanoscience Delft University of Technology Delft 2628 CD NL

Department of Imaging Physics Delft University of Technology Delft 2628 CJ NL

Institute of Macromolecular Chemistry Czech Academy of Sciences Prague 162 00

Institute of Scientific Instruments Czech Academy of Sciences Brno 61264

Kavli Institute of Nanoscience Delft University of Technology Delft 2628 CJ NL

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