Related Experiment Video
Updated: Jan 28, 2026
09:09
Preparation and Delivery of Protein Microcrystals in Lipidic Cubic Phase for Serial Femtosecond Crystallography
Published on: September 20, 2016
11.9K
Automated microbatch-under-oil phase diagrams to rationalize serial crystallography sample preparation
Jack Stubbs1, Courtney J Tremlett2, Abigail Waitman1
1School of Biological Sciences, Faculty of Environmental and Life Sciences, University of Southampton, Southampton SO17 1BJ, United Kingdom.
Iucrj
|January 27, 2026
Summary
This study introduces an automated microbatch-under-oil method for rapid protein crystallization screening, significantly reducing sample needs for serial crystallography. The technique optimizes microcrystal production for advanced diffraction experiments.
Area of Science:
- Structural Biology
- Biochemistry
- Crystallography
Background:
- Serial crystallography requires high-density microcrystal suspensions.
- Sample optimization for microcrystal production is a bottleneck.
- Traditional phase diagram mapping consumes excessive protein sample.
Purpose of the Study:
- To develop a rapid, low-volume method for mapping protein crystallization phase boundaries.
- To optimize microcrystal density, size, and lattice order for serial crystallography.
- To provide a reproducible workflow for both known and new protein targets.
Main Methods:
- Automated microbatch-under-oil crystallization.
- Diagonal sampling strategy varying protein-to-precipitant ratios.
- Linked variation of multiple precipitants and automated seed-stock titration.
Main Results:
- Rapid phase boundary mapping using minimal protein (15-60 µl).
- Identification of nucleation zones and morphology-specific regions.
- Precise definition of the metastable zone for nucleation-limited systems.
Conclusions:
- The integrated approach systematically decouples nucleation from growth.
- Ensures stable chemical coordinates for scale-up, unlike vapor diffusion.
- Transforms empirical sample preparation into a rational, efficient process for serial diffraction.
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