Thermo Fisher Scientific 1 article published in JoVE Biochemistry Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope Adrian Koh*1, Sagar Khavnekar*2, Wen Yang1, Dimple Karia1, Dennis Cats1, Rob van der Ploeg1, Fanis Grollios1, Oliver Raschdorf1, Abhay Kotecha1, Daniel Němeček3 1Materials and Structural Analysis Division, Thermo Fisher Scientific, 2Max Planck Institute of Biochemistry, Molecular Structural Biology, 3Materials and Structural Analysis Division, Thermo Fisher Scientific High-resolution cryo-EM maps of macromolecules can be also achieved by using 200 kV TEM microscopes. This protocol shows the best practices for setting accurate optics alignments, data acquisition schemes, and selection of imaging areas that are all essential for the successful collection of high-resolution datasets using a 200 kV TEM.