ID23-2 Summary

ID23-2 operates at a fixed energy and is in particular specialized to perform experiments with very small protein crystals and for serial crystallography.

ID23-2 delivers a flux of about 1x1013 ph/s in a microfocus beam. The beam is typically detuned to a size of ~4x4 µm FWHM. Since the recent upgrade, the beam is vertically focused by CRLs and horizontally focused by a mirror in half-Kirkpatrick-Baez (KB) geometry. The beamline is equipped with a MD3-UP microdiffractometer with a vertical spindle that can perform very fast mesh scans and is compatible with in-situ data collection for low-profile crystallization plates. A FlexHCD sample changer robot has been installed that can take up to 23 Unipucks (NEITHER SPINE Pucks/SC3 Baskets nor non-SPINE pins can be used on this beamline), please see sample changer section for details). The detector is an Eiger2 X 9m

Video Synopsis of usage:

Recent News

Having problems with small crystals or plates rotating out of the beam? There is a new workflow to recalibrate the rotation axis diffractively, 

January 2023: The new Eiger2 X 9M detector has been installed and commissioned

March 2022: The beamline rebuild paper has been published:

 ID23-2: an automated and high-performance microfocus beamline for macromolecular crystallography at the ESRF
Nanao, M., Basu, S., Zander, U., Giraud, T., Surr, J., Guijarro, M., Lentini, M., Felisaz, F., Sinoir, J., Morawe, C., Vivo, A., Beteva, A., Oscarsson, M., Caserotto, H., Dobias, F., Flot, D., Nurizzo, D., Gigmes, J., Foos, N., Siebrecht, R., Roth, T., Theveneau, P., Svensson, O., Papp, G., Lavault, B., Cipriani, F., Barrett, R., Clavel, C. & Leonard, G. (2022). J. Synchrotron Rad. 29, 581--590.

November 2021: The sybil control computer has been replaced with a new machine called "id232control"

October 2020: the Kappa has been dismounted for repair

Remote acces information here: https://www.esrf.eu/UsersAndScience/Experiments/MX/How_to_use_our_beamlines/remote-access

 

 

Scientific Applications

Protein crystal samples are becoming ever smaller, with today's samples typically around 10 to 100 microns in size. With the development of third generation synchrotron radiation sources, it is now possible to have access to a focused and very intense hard X-ray beam in the µm-range. ID23-2 produces a focused beam of less than 5 µm in diameter (FWHM) to allow to nable the study of microcrystals as well as facilitate serial crystallographic experiments.

relevant publications:

Small is beautiful: protein micro-crystallography ; Cusack S, Belrhali H, Bram A, et al.  Nature Structural Biology 5: 634-637 Suppl. S AUG 1998

Protein microcrystals and the design of a microdiffractometer: current experience and plans at EMBL and ESRF/ID13 ; Perrakis A, Cipriani F, Castagna JC, et al.  Acta Crystallographica Section D-Biological Crystallography 55: 1765-1770 Part 10 OCT 1999

ID23-2: an automated and high-performance microfocus beamline for macromolecular crystallography at the ESRF
Nanao, M., Basu, S., Zander, U., Giraud, T., Surr, J., Guijarro, M., Lentini, M., Felisaz, F., Sinoir, J., Morawe, C., Vivo, A., Beteva, A., Oscarsson, M., Caserotto, H., Dobias, F., Flot, D., Nurizzo, D., Gigmes, J., Foos, N., Siebrecht, R., Roth, T., Theveneau, P., Svensson, O., Papp, G., Lavault, B., Cipriani, F., Barrett, R., Clavel, C. & Leonard, G. (2022). J. Synchrotron Rad. 29, 581--590.

Techniques Available

  • Microfocus beam
  • Fixed wavelength rapid data collection (SAD/MIR)
  • Fast mesh scans
  • In-situ data collection available upon request
  • HC1 humidity control device available upon request

Warning

When coming on ID23-2 as a user, you will have to deal with a small and intense beam. Keeping a 2000 µm3 (about 20 x 10 x 10 µm3) crystal in a 5 µm diameter beam needs good beam stability AND accurate crystal centring.

To maximise success rates and data quality, we recommend using the minimum amount of cryoprotection possible (or no cryoprotectant at all).  This can be important for reducing background as well as optical centering of crystals, although X-ray centering is also available on the beamline.

Beam drifts

In normal operating conditions, small beam drifts (variations in position of less than 10 µm in both vertically and horizontally directions) have been regularly observed over a period of 12 hours. Such variations are expected and are mainly driven by experimental hutch temperature variations. In order to minimize those it is highly advisable to enter the control cabin with as few persons as possible at the same time and to open both the door of the experimental hutch as well as the door outside the experimental hutch only when necessary.

The possibility of beam drift makes regular beam position checks necessary. This can simply be done by using the center beam button in MXCube.

 

EMBL has provided a scientist position to operate the ID23-2 beamline.