JADAS: a customizable automated data acquisition system and its application to ice-embedded single particles

Area of Science:

• Microscopy
• Structural Biology
• Biophysics

Background:

• Transmission electron microscopy (TEM) is crucial for visualizing biological macromolecules.
• Automated data acquisition is essential for efficient high-resolution structure determination.
• JEOL Transmission Electron Microscopes offer advanced capabilities for cryo-electron microscopy.

Purpose of the Study:

• To introduce and describe the JEOL Automated Data Acquisition System (JADAS).
• To demonstrate JADAS's capability in automating image acquisition for ice-embedded single particles.
• To validate JADAS performance by achieving subnanometer resolution structure of bacteriophage epsilon 15.

Main Methods:

• Utilized JADAS software for automated image acquisition on JEOL Transmission Electron Microscopes.
• Employed low-dose imaging conditions suitable for sensitive biological samples.
• Customized imaging operation sequences for optimal data collection.
• Applied JADAS to image ice-embedded bacteriophage epsilon 15 using a JEM3200FSC electron cryo-microscope.

Main Results:

• JADAS accurately locates and images suitable specimen areas on cryo-EM grids.
• Sufficient high-quality images for structural analysis were collected within an 8-hour session.
• A subnanometer resolution structure of bacteriophage epsilon 15 was obtained.
• The determined structure is in agreement with previously reported results.

Conclusions:

• JADAS significantly enhances the efficiency of single-particle cryo-electron microscopy data acquisition.
• The software provides flexibility for user customization in imaging protocols.
• JADAS facilitates rapid structure determination of biological macromolecules.