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.
Highlights
This paper focuses on using vapour diffusion tools to make the conversion into batch as these are generally more widely used than microbatch, but the conversion could be made using microbatch techniques instead (Chayen, 1998)
The aim of this paper was to suggest methods and ideas to aid in converting a vapour diffusion crystallization experiment into a larger-scale batch experiment
Vapour diffusion crystallization experiments can be converted into batch crystallization by understanding the role the precipitant is playing in the crystallization process and looking at the timescale of crystal nucleation and growth
Summary
For certain types of protein crystal, those of viral capsid proteins, cryo-cooling is not possible and the merging of multiple small wedge rotations is a necessary and effective way of acquiring a complete data set (Fry et al, 1999). The XFEL beam destroys the sample upon interaction (Neutze et al, 2000), precluding wedged data collection, and takes serial data collection to its logical extreme, i.e. one image per crystal. This necessitates the need for the delivery of a steady stream of hundreds or thousands of micro-crystals into the path of the X-ray beam in order to sample reciprocal space appropriately
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