Abstract

An approach for serial crystallography experiments based on wedged-data collection is described. This is an alternative method for recording in situ X-ray diffraction data on crystalline samples efficiently loaded in an X-ray compatible microfluidic chip. Proper handling of the microfluidic chip places crystalline samples at geometrically known positions with respect to the focused X-ray interaction area for serial data collection of small wedges. The integration of this strategy takes advantage of the greatly modular sample environment available on the endstation, which allows access to both in situ and more classical cryo-crystallography with minimum time loss. The method represents another optional data collection approach that adds up to the already large set of methods made available to users. Coupled with the advances in processing serial crystallography data, the wedged-data collection strategy proves highly efficient in minimizing the amount of required sample crystals for recording a complete dataset. From the advances in microfluidic technology presented here, high-throughput room-temperature crystallography experiments may become routine and should be easily extended to industrial use.

Highlights

  • Serial protein crystallography at synchrotrons has grown in interest following the developments at X-ray free-electron laser (XFEL) sources for X-ray diffraction data collection (Grunbein & Kovacs, 2019; Chavas et al, 2015) with reduced damages (Nass, 2019), a serious obstacle for sensitive biological samples

  • In the course of this work, the microfluidic chips developed for the In situ geometrically optimized raster (IGOR) scanning have been subject to constant improvement

  • The exciting progress witnessed in the development of microfluidic chips applied to biological objects illustrates the great potential of these devices for handling samples that are classically difficult to manipulate, fragile or even hazardous

Read more

Summary

Introduction

Serial protein crystallography at synchrotrons has grown in interest following the developments at X-ray free-electron laser (XFEL) sources for X-ray diffraction data collection (Grunbein & Kovacs, 2019; Chavas et al, 2015) with reduced damages (Nass, 2019), a serious obstacle for sensitive biological samples. For the XFEL approach, in contrast to the more classical synchrotron cryo-crystallography, hundreds of thousands of in situ crystals are exposed to extremely intense X-rays that lead to a unique diffraction image recorded per crystal using the principle referred to as ‘diffraction before destruction’ (Neutze et al, 2000; Johansson et al, 2017). XFEL and synchrotron sources differ in the nature of the photons being distributed, which, in turn, affects the type of studies that can be performed, the overall instrumentation on the experimental station, how the sample should be prepared and handled, and how data analysis needs to be carried out.

Methods
Results
Conclusion
Full Text
Published version (Free)

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 call