Comprehensive microcrystal electron diffraction sample preparation for cryo-EM

Area of Science:

• Structural biology
• Biophysics
• Materials science

Background:

• Microcrystal Electron Diffraction (MicroED) is a powerful cryogenic electron microscopy (cryo-EM) technique for determining crystal structures.
• It is applicable to a wide range of samples, including small molecules, soluble proteins, and membrane proteins.
• Expertise in MicroED sample preparation, data acquisition, and processing is not always readily available.

Purpose of the Study:

• To provide a comprehensive guide on MicroED sample preparation.
• To address commonly used methods for various sample categories.
• To offer instructions and optimization strategies for MicroED sample preparation.

Main Methods:

• Detailed protocols for MicroED sample preparation are presented.
• Methods cover room temperature solid-state small molecules, soluble proteins, and membrane protein crystals.
• Optimization strategies are included for unique crystal growth and preparation conditions.

Main Results:

• The guide offers practical instructions for new users of MicroED.
• It provides a framework for optimizing sample preparation for diverse crystalline materials.
• The protocol is suitable for researchers with existing expertise in biochemistry, crystallography, and cryo-EM.

Conclusions:

• This resource aims to improve accessibility to MicroED technology.
• Effective sample preparation is crucial for successful MicroED structure determination.
• The guide empowers researchers to overcome challenges in MicroED sample preparation.