Preparing Frozen-Hydrated Protein–Nucleic Acid Assemblies for High-Resolution Cryo-EM Imaging
Methods in molecular biology, January 2018
Panchali Goswami, Julia Locke, Alessandro Costa
High-resolution image acquisition and structure determination by cryo-electron microscopy is becoming increasingly streamlined. Preparing electron-microscopy grids of suitable quality remains, however, a critical bottleneck. Strategies to achieve particle monodispersity, optimal sample concentration and suitable ice thickness can vary from specimen to specimen. In this book chapter we describe our protocols for negative-stain grid and cryo-grid preparation, which we apply to studying protein-nucleic acid complexes.
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