Protocol for the use of focused ion-beam milling to prepare crystalline lamellae for microcrystal electron diffraction (MicroED)
Michael W. Martynowycz, Tamir Gonen
Abstract
We present an in-depth protocol to reproducibly prepare crystalline lamellae from protein crystals for subsequent microcrystal electron diffraction (MicroED) experiments. This protocol covers typical soluble proteins and membrane proteins embedded in dense media. Following these steps will allow the user to prepare crystalline lamellae for protein structure determination by MicroED. For complete details on the use and execution of this protocol, please refer to Martynowycz et al. (2019a, 2020a).
Topics & Concepts
Electron diffractionDiffractionElectron crystallographyNanotechnologyMaterials scienceCrystallographyChemistryOpticsPhysicsEnzyme Structure and FunctionPorphyrin Metabolism and DisordersBiochemical and Molecular Research