Understanding the invisible hands of sample preparation for cryo-EM

Cryo-electron microscopy (cryo-EM) is rapidly becoming an attractive method in the field of structural biology. With the exploding popularity of cryo-EM, sample preparation must evolve to prevent congestion in the workflow. The dire need for improved microscopy samples has led to a diversification of methods. This Review aims to categorize and explain the principles behind various techniques in the preparation of vitrified samples for the electron microscope. Various aspects and challenges in the workflow are discussed, from sample optimization and carriers to deposition and vitrification. Reliable and versatile specimen preparation remains a challenge, and we hope to give guidelines and posit future directions for improvement.

Automated summary

Cryo-electron microscopy is gaining popularity in structural biology, leading to the need for advanced sample preparation techniques to optimize workflow efficiency. This review categorizes and explains various methods for preparing vitrified samples for electron microscopy, discussing challenges and future directions for improvement. Reliable and versatile specimen preparation remains a key challenge in the field, with guidelines provided to address these issues.