1. SFX and Laue Diffraction
Quan Hao
School of Biomedical Sciences
12/2/2018

3. Diffraction before Destruction Sub-micron Crystals Room Temperature
Serial Femtosecond Crystallography (SFX)：
Diffraction before Destruction
Sub-micron Crystals
Room Temperature

4. APS data (7.7 Å) vs. LCLS data (3.3 Å)
Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser. Nature 523, 561 (2015).
APS data (7.7 Å) vs. LCLS data (3.3 Å)
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5. Structural basis for selectivity and diversity in angiotensin II receptors Haitao Zhang et al. Nature 544, 327–332 (20 April 2017)
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6. Zhang at al. Volume 2 | Part 3 | May 2015 | Pages 322–326
A one-dimensional powder diffraction pattern (e.g. from a mixture of zeolites) seen using conventional methods (left) may potentially be analysed as three-dimensional single-crystal patterns using serial crystallography (right).
A one-dimensional powder diffraction pattern seen using conventional methods (left) may potentially be analysed as three-dimensional single-crystal patterns using serial crystallography (right). [Hao, Q. (2015). IUCrJ, 2, doi: /S ]
Zhang at al. Volume 2 | Part 3 | May 2015 | Pages 322–326
Hao, Volume 2 | Part 3 | May 2015 | Pages 307–308

7. Improve SFX?
Challenges: partial reflections; massive number of crystals (~1M) needed; difficult to process.
Proposed SwissFEL bandwidth ~ 3% by strong pulse compression (save time and sample by 30-fold comparing to mono mode)
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8. Laue Diffraction: a stationary crystal during exposure; polychromatic beam; wavelength normalization required.
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9. Laue Diffraction Large number of spots on a single image.
Non-oscillation sample (e.g. in situ plates)
Potential time-resolved application
Examples:
Moffat, Szebenyi & Bilderback (1984), Science, 223,
Hadju et al (1987) Nature, 329,
Bilderback Hoffman & Thiel (1994) Science, 263,
Genick, …, Moffat & Getzoff (1997) Science, 275,
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10. Laue v.s. Mono
Laue
Pro: most full reflections; fewer crystals needed; easier to scale/merge.
Con: more sensitive to mosaicity; possible spatial overlap for large unit cell.
Mono
Pro: less sensitive to mosaicity.
Con: partial reflections; more crystals needed; difficult to process.
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12. Experimental Procedures
MicroMesh (Mitegen) mounts and flash-cooled, or
Liquid jet or LCP or in situ plates at room-T
Data will be recorded from one crystal at a time using an area detector. For each crystal, one still image will be recorded
Data will be indexed, refined and integrated with the program LAUEGEN
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13. Feasibility study @CHESS-D1 Cornaby,S. , D. M. E. Szebenyi, D. -M
Feasibility Cornaby,S., D. M. E. Szebenyi, D.-M. Smilgies, D. J. Schuller, R. Gillilan, Q. Hao and D. H. Bilderback (2010) Acta Cryst. D66, 2-11.
ΔE/E ~30%
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14. Narrow bandpass Optics
Kazmirov et al (2006) J. Synchrotron Rad. 13, Multilayer X-ray optics at CHESS:
ΔE/E: 0.2%, 2%, 5% and 10%
W/B4C: most commonly used
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15. CHESS-G1 experiment Multi-layer optic (~1.7% bandpass: 0.975-0.992 Å)
One set of slits (~0.1 x 0.1 mm2)
Lysozyme crystal (a=b=79, c=38Å, P43212)
Room temperature
72 still images; =2.5; 1 s exposure
Crystal-detector (Q4) dist. = 188 mm
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16. Diffraction image
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17. Data processing Using DENZO/MOSFLM to find crystal orientation
Using LAUEGEN* to refine orientation, improve soft limits (lmin,lmax & dmin) and integrate
Using LAUENORM to normalize, scale and merge.
*Campbell, J. W., Hao, Q., Harding, M. M., Nguti, N. D. & Wilkinson, C. (1998). LAUEGEN version 6.0 and INTLDM. J. Appl. Cryst. 31,
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19. Data statistics Resolution range: 12.0 – 2.2 Å
Unique reflections: 4370
I/(I) = 10.0
Rmerge = 13.4% (~6% between Å)
Redundancy = 2.0
Completeness = 70.7%
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20. Structure solution and refinement
Straightforward Molecular replacement solution with AMoRe: c.c.=75%, R=31.7%
Refinement with REFMAC: R ~ 20%.
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21. Electron density map (residues 40-45, 50-55, 58-60)
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28. Discussion Ideal bandwidth? small unit cells (<100Å), ~10%;
large unit cells (>100Å); ~6%.
At 1.7% bandwidth, ~1°.
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29. Proposed feasibility study at SSRF
Beamline: a PX beamline with Multilayer-optics
Scientific Goal：
To determine protein structure using sub-micron crystals
Experimental：
Laue diffraction; One-shot-per-crystal structure determination
Specifications：
Photon energy： 7-16 keV
Bandwidth：>3%
Beamsize：～20μm x 20μm
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30. Computation – future plan
Daresbury Laue Software Suite
LAUEGEN: new algorithm to find orientation;
multi-core code to process large amount of images.
LAUENORM: better curve-fitting for narrow bandwidth.
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31. Acknowledgements MacCHESS:
Qun Liu, Qingqiu Huang, Chris Heaton, Bill Miller, Marian Szebenyi.
CHESS:
Detlef Smilgies, Joel Brock, Ernie Fontes, Sterling Cornaby, Don Bilderback, Sol Gruner.
Daresbury Lab:
John Helliwell, John Campbell, Marjorie Harding.
Institute of Physics, CAS
Haifu Fan, Tao Zhang
Institute of Applied Physics, CAS
Dong Wang, Haixiao Deng
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