3D scanning electron microscopy has become an important tool to answer scientific questions in life science. The aim of the course is to provide hands-on experience concerning sample preparation for the main serial imaging techniques in scanning electron microscopy (SEM): microtome-based serial block-face SEM, focused ion beam SEM and array tomography. Chemical fixation (bench processing and microwave-assisted processing) as well as high-pressure freezing and freeze substitution for scanning electron microscopy will be performed. The combination of theoretical and practical sessions will give the participants the possibility to assess the potentials of each sample preparation technique and decide which are most suitable for their own scientific project and research question. The course will also cover data processing and analysis, as these tools are essential to all 3D electron microscopy techniques.