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Successful sample preparation for serial crystallography experiments

DOI: 10.1107/S1600576719013517 DOI Help

Authors: John H. Beale (Diamond Light Source) , Rachel Bolton (Diamond Light Source; University of Southampton Centre for Biological Sciences) , Stephen A. Marshall (The University of Manchester) , Emma V. Beale (Diamond Light Source) , Stephen B. Carr (Research Complex at Harwell) , Ali Ebrahim (Diamond Light Source; University of Essex) , Tadeo Moreno-chicano (Institute de Biologie Structurale) , Michael A. Hough (University of Essex) , Jonathan A. R. Worrall (University of Essex) , Ivo Tews (University of Southampton) , Robin L. Owen (Diamond Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Applied Crystallography , VOL 52 , PAGES 1385 - 1396

State: Published (Approved)
Published: December 2019

Open Access Open Access

Abstract: Serial crystallography, at both synchrotron and X-ray free-electron laser light sources, is becoming increasingly popular. However, the tools in the majority of crystallization laboratories are focused on producing large single crystals by vapour diffusion that fit the cryo-cooled paradigm of modern synchrotron crystallography. This paper presents several case studies and some ideas and strategies on how to perform the conversion from a single crystal grown by vapour diffusion to the many thousands of micro-crystals required for modern serial crystallography grown by batch crystallization. These case studies aim to show (i) how vapour diffusion conditions can be converted into batch by optimizing the length of time crystals take to appear; (ii) how an understanding of the crystallization phase diagram can act as a guide when designing batch crystallization protocols; and (iii) an accessible methodology when attempting to scale batch conditions to larger volumes. These methods are needed to minimize the sample preparation gap between standard rotation crystallography and dedicated serial laboratories, ultimately making serial crystallography more accessible to all crystallographers.

Journal Keywords: serial macromolecular crystallography; XFELs; batch crystallization; vapour diffusion; micro-crystallization

Subject Areas: Technique Development


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