Novel combined crystallization plate for high-throughput crystal screening and in situ data collection at a crystallography beamline

Liang, M.; Yu, L.; Wang, Z.; Zhou, H.; Zhang, Y.; Wang, Q.; He, J.

doi:10.1107/S2053230X21008104

Novel combined crystallization plate for high-throughput crystal screening and in situ data collection at a crystallography beamline

M. Liang, L. Yu, Z. Wang, H. Zhou, Y. Zhang, Q. Wang and J. He

In situ microplates are small in size, crystal cultivation and operation are difficult, and the efficiency of crystal screening is relatively low. To solve this problem, a novel combined crystallization plate was designed for high-throughput crystal cultivation and in situ data collection. A frame was used to hold 48 in situ microplates, and the in situ microplates were sealed on one side with an ultralow background-scattering Kapton film. An automatic liquid handler (Mosquito) was used to add a liquid drop to the in situ microplates in the frame, and CrystalClear HD tape was used to seal the frame. A sealed frame holding 48 microplates was developed as a novel combined crystallization plate and was used for crystal cultivation under different conditions and in situ data collection at the synchrotron beamline. Moreover, individual microplates can be separated from the combined crystal plate and then fixed on a magnetic base or loaded onto a UniPuck for in situ data collection. Automatic grid scanning was used to locate crystals. The efficiency of the combined crystallization plate for crystal screening was verified. This method avoids the manual manipulation of crystals during crystal screening and diffraction data collection; therefore, the combined crystallization plate is suitable for large-scale screening of microcrystals.

Keywords: in situ diffraction; automatic sampling; automatic sample loading; crystal screening; crystallization plates.

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Supporting information

	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup1.mpg Supplementary Movie S1. The in situ microplate is placed on the supporting frame.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup2.mpg Supplementary Movie S2. The in situ microplates are single side sealed on the supporting frame.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup3.mpg Supplementary Movie S3. Cut along the cutting grooves of the supporting frame with a blade to obtain single side sealed in situ microplate.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup4.mpg Supplementary Movie S4. The dam bar is placed in the strip groove of the holding frame.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup5.mpg Supplementary Movie S5. Put in situ microplates into the small grooves of the supporting frame.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup6.mpg Supplementary Movie S6. The automatic liquid handler Mosquito was used to automatically add samples to the spotting holes.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup7.mpg Supplementary Movie S7. Scotch tape is used to seal the holding frame to generate a new combinated crystallography plate.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup8.mpg Supplementary Movie S8. The dam bar was removed.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup9.mpg Supplementary Movie S9. In situ microplates will be pushed into the large groove of the supporting frame.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup10.mpg Supplementary Movie S10. The in situ microplate was sealed by a film with low background scattering properties such as Kapton film.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup11.mpg Supplementary Movie S11. A blade is used to cut along the large groove of the supporting frame.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup12.mpg Supplementary Movie S12. The in situ microplate is contained in the crystal sample box UniPuck.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup13.mpg Supplementary Movie S13. 3D models of the microplate, holding frame, supporting frame and dam bar.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup14.mpg Supplementary Movie S14. Tweezers are used to take out the single in situ microplate and coat it individually on the supporting frame.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup15.mpg Supplementary Movie S15. One column in situ microplates were taken out for coating.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup16.mpg Supplementary Movie S16. Manual screening of the crystals.
	Moving Picture Experts Group (MPG) video file https://doi.org/10.1107/S2053230X21008104/nj5308sup17.mpg Supplementary Movie S17. Automatic screening of the crystals.

PDB reference: lysozyme, 7f26

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Format		BIBTeX
		EndNote
		RefMan
		Refer
		Medline
		CIF
		SGML
		Text
		Plain Text

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