Abstract
Synchrotron serial crystallography (SSX) is an emerging data-collection method for micro-crystallography on synchrotron macromolecular (MX) crystallography beamlines. At SPring-8, the feasibility of the fixed-target approach was examined by collecting data using a 2D raster scan combined with goniometer rotation. Results at cryogenic temperatures demonstrated that rotation is effective for efficient data collection in SSX and the method was named serial synchrotron rotation crystallography (SS-ROX). To use this method for room-temperature (RT) data collection, a humid air and glue-coating (HAG) method was developed in which data were collected from polyvinyl alcohol-coated microcrystals fixed on a loop under humidity-controlled air. The performance and the RT data-collection strategy for micro-crystallography were evaluated using microcrystals of lysozyme. Although a change in unit-cell dimensions of up to 1% was observed during data collection, the impact on data quality was marginal. A comparison of data obtained at various absorbed doses revealed that absorbed doses of up to 210 kGy were tolerable in both global and local damage. Although this limits the number of photons deposited on each crystal, increasing the number of merged images improved the resolution. On the basis of these results, an equation was proposed that relates the achievable resolution to the total photon flux used to obtain a data set.
Highlights
Cryocrystallography is an essential technique to maximize the benefits of the high-brilliance beams available at synchrotron facilities by mitigating radiation damage (Garman & Owen, 2006)
The importance of structure determination at room temperature (RT) has been rediscovered in recent years
We have demonstrated that humid air and glue-coating (HAG) SS-ROX is appliable to RT micro-crystallography and have evaluated an RT data-collection strategy
Summary
Cryocrystallography is an essential technique to maximize the benefits of the high-brilliance beams available at synchrotron facilities by mitigating radiation damage (Garman & Owen, 2006). In the case of CT, a useful data-collection method is multiple small-wedge data collection, in which data are collected from each crystal using an angular range of a few degrees to 10 and are merged after data collection. By combining this method with a 2D raster scan to identify the position of the crystal on the diffraction base, data collection can be automated. This makes it easy to use hundreds of crystals or more to complete the data set (Hirata et al, 2019). An alternative efficient method is needed for RT data collection because the number of crystals needed to complete the data set is larger and the 2D raster scan prior to data collection needs to be avoided
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Acta crystallographica. Section D, Structural biology
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
