Kavli Affiliate: Michael Beer
| Authors: Jos J. A. G. Kamps, Philip Hinchliffe, Johan Glerup, Emily I. Freeman, Pauline A. Lang, Catherine L Tooke, Michael Beer, Laura Parkinson, Do-Heon Gu, Sehan Park, Nicholas Devenish, Tiankun Zhou, Anastasya Shilova, Patrick Rabe, Christopher J Schofield, Jim Spencer, Jaehyun Park, Robin Owen, Allen M. Orville and Pierre Aller
| Summary:
We describe the design and implementation of a drop on fixed target method for time-resolved serial crystallography at both synchrotron and XFEL facilities. A piezoelectric droplet dispensing pipette is employed for addition of picolitre volume (40 – 90 pL) aqueous droplets, containing (co-)substrate(s) or ligand(s), onto enzyme microcrystals immobilised on a solid support. The system was tested with various enzyme systems, including lysozyme and two ꞵ-lactamases, CTX-M-15 and AmpCEC. Mitigation strategies for cross-well contamination, including the implementation of interleaved controls, are described; the overall performance of the system at synchrotron and X-ray free electron laser facilities was evaluated. This drop on fixed target method is a reliable framework for time-resolved crystallography and will improve the consistency of measurements across facilities.