Droplet microfluidics for time-resolved serial crystallography

DOI: 10.1107/S2052252524001799

Authors: Jack Stubbs (University of Southampton; Diamond Light Source) , Theo Hornsey (University of Southampton) , Niall Hanrahan (University of Southampton) , Luis Blay Esteban (Universitat Carlemany) , Rachel Bolton (University of Southampton; Diamond Light Source) , Martin Maly (University of Southampton) , Shibom Basu (European Molecular Biology Laboratory) , Julien Orlans (European Synchrotron Radiation Facility (ESRF)) , Daniele De Sanctis (European Synchrotron Radiation Facility (ESRF)) , Jung-Uk Shim (University of Leeds) , Patrick D. Shaw Stewart (Douglas Instruments Ltd) , Allen M. Orville (Diamond Light Source; Research Complex at Harwell) , Ivo Tews (University of Southampton) , Jonathan West (University of Southampton)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Iucrj , VOL 11 , PAGES 237 - 248

State: Published (Approved)
Published: March 2024

Abstract: Serial crystallography requires large numbers of microcrystals and robust strategies to rapidly apply substrates to initiate reactions in time-resolved studies. Here, we report the use of droplet miniaturization for the controlled production of uniform crystals, providing an avenue for controlled substrate addition and synchronous reaction initiation. The approach was evaluated using two enzymatic systems, yielding 3 µm crystals of lysozyme and 2 µm crystals of Pdx1, an Arabidopsis enzyme involved in vitamin B6 biosynthesis. A seeding strategy was used to overcome the improbability of Pdx1 nucleation occurring with diminishing droplet volumes. Convection within droplets was exploited for rapid crystal mixing with ligands. Mixing times of <2 ms were achieved. Droplet microfluidics for crystal size engineering and rapid micromixing can be utilized to advance time-resolved serial crystallography.

Journal Keywords: droplet microfluidics; crystal miniaturization; micromixing; time-resolved serial crystallography

Diamond Keywords: Enzymes

Subject Areas: Technique Development, Biology and Bio-materials

Diamond Offline Facilities: XFEL-Hub

Facility: ID29 at ESRF

Added On: 11/03/2024 11:30

Documents:
zf5024.pdf

Discipline Tags:

Technique Development - Life Sciences & Biotech Life Sciences & Biotech

Technical Tags: