Download citation
Download citation
link to html
Serial femtosecond crystallography (SFX) enables the determination of room-temperature crystal structures of macromolecules with minimized radiation damage and provides time-resolved molecular dynamics by pump–probe or mix-and-inject experiments. In SFX, a variety of sample delivery methods with unique advantages have been developed and applied. The combination of existing sample delivery methods can enable a new approach to SFX data collection that combines the advantages of the individual methods. This study introduces a combined inject-and-transfer system (BITS) method for sample delivery in SFX experiments: a hybrid injection and fixed-target scanning method. BITS allows for solution samples to be reliably deposited on ultraviolet ozone (UVO)-treated polyimide films, at a minimum flow rate of 0.5 nl min−1, in both vertical and horizontal scanning modes. To utilize BITS in SFX experiments, lysozyme crystal samples were embedded in a viscous lard medium and injected at flow rates of 50–100 nl min−1 through a syringe needle onto a UVO-treated polyimide film, which was mounted on a fixed-target scan stage. The crystal samples deposited on the film were raster scanned with an X-ray free electron laser using a motion stage in both horizontal and vertical directions. Using the BITS method, the room-temperature structure of lysozyme was successfully determined at a resolution of 2.1 Å, and thus BITS could be utilized in future SFX experiments.

Supporting information

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup1.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup2.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup3.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup4.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup5.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup6.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup7.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup8.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup9.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup10.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup11.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup12.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup13.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup14.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup15.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup16.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup17.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup18.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup19.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup20.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup21.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup22.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup23.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup24.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup25.mp4
Supplementary material

mp4

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S1600576722005556/te5095sup26.mp4
Supplementary material

pdf

Portable Document Format (PDF) file https://doi.org/10.1107/S1600576722005556/te5095sup27.pdf
Supporting figures and BITS code

PDB references: 7wkr; 7wuc


Follow J. Appl. Cryst.
Sign up for e-alerts
Follow J. Appl. Cryst. on Twitter
Follow us on facebook
Sign up for RSS feeds