Automated grid handling and image acquisition for two-dimensional crystal screening

Area of Science:

• Structural biology
• Biophysics
• Macromolecular crystallography

Background:

• Two-dimensional (2D) crystallization is crucial for determining protein structures.
• Large-scale crystallization trials generate numerous samples requiring efficient imaging.
• Current imaging protocols can be time-consuming and labor-intensive.

Purpose of the Study:

• To describe an automated imaging protocol for large-scale 2D crystallization trials.
• To evaluate the efficiency of a two-pass imaging strategy using Leginon and a robotic grid handler.
• To explore the potential for fully automated target selection in 2D crystallization imaging.

Main Methods:

• Implementation of a two-pass imaging protocol.
• Utilizing Leginon software for automated microscopy control.
• Employing a grid handling robot for sample manipulation.
• Manual target selection to bridge imaging passes.

Main Results:

• The described protocol effectively images a large number of grids from 2D crystallization trials.
• The two-pass imaging approach, combined with manual target selection, is functional.
• Potential exists for combining the two passes through automated identification of targets at various trial stages.

Conclusions:

• Automation of large-scale 2D crystallization imaging is achievable.
• The described protocol offers a significant improvement in efficiency for structural biology workflows.
• Further development in automated target recognition could lead to fully automated imaging solutions.