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Acknowledgements

Acknowledgements. Ken Frankel Chris Neilson Michael Blum Joe Ferrara. 8.3.1 creator: Tom Alber 8.3.1 PRT head: Jamie Cate Center for Structure of Membrane Proteins Membrane Protein Expression Center II Center for HIV Accessory and Regulatory Complexes W. M. Keck Foundation

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Acknowledgements

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  1. Acknowledgements Ken Frankel Chris Neilson Michael Blum Joe Ferrara 8.3.1 creator: Tom Alber 8.3.1 PRT head: Jamie Cate Center for Structure of Membrane Proteins Membrane Protein Expression Center II Center for HIV Accessory and Regulatory Complexes W. M. Keck Foundation Plexxikon, Inc. M D Anderson CRC University of California Berkeley University of California San Francisco National Science Foundation University of California Campus-Laboratory Collaboration Grant Henry Wheeler The Advanced Light Source is supported by the Director, Office of Science, Office of Basic Energy Sciences, Materials Sciences Division, of the US Department of Energy under contract No. DE-AC02-05CH11231 at Lawrence Berkeley National Laboratory.

  2. The “success rate” of structure determination 100 s/dataset 200 days/year ~150 beamlines ~26,000,000 datasets/year 8000 PDBs in 2010

  3. DVD data archive ~2500 datasets/year ~100 PDBs/year

  4. Why do structures fail? Overlaps Radiation Damage Signal to noise

  5. Why do structures fail? Overlaps Radiation Damage Signal to noise

  6. Why do structures fail? Overlaps Radiation Damage Signal to noise

  7. Why do structures fail? Overlaps Radiation Damage Signal to noise

  8. signalvsnoise

  9. signalvsnoise

  10. signalvsnoise “If you don’t have good data,then you have no data at all.” -Sung-Hou Kim

  11. signalvsnoise easy hard impossible

  12. signalvsnoise easy hard impossible threshold of “solvability”

  13. signalvsnoise “If you don’t have good data,then you must learn statistics.” -James Holton

  14. Adding noise

  15. Adding noise 12 + 12 = 1.42

  16. Adding noise 12 + 12 = 1.42 32 + 12 = 3.22 σtotal2 = σ12 + σ22

  17. Adding noise 12 + 12 = 1.42 32 + 12 = 3.22 σtotal2 = σ12 + σ22

  18. Adding noise 12 + 12 = 1.42 32 + 12 = 3.22 σtotal2 = σ12 + σ22

  19. Adding noise 12 + 12 = 1.42 32 + 12 = 3.22 102 + 12 = 10.052

  20. “photon counting” Read-out noise Shutter jitter Beam flicker spot shape radiation damage σ(N) = sqrt(N) rms 11.5 e-/pixel rms 0.57 ms 0.15 %/√Hz pixels? mosaicity? B/Gray? signalvsnoise

  21. Simulated diffraction imageMLFSOM simulated real

  22. mosaic spread = 0 º

  23. mosaic spread = 0.1º

  24. mosaic spread = 0.2º

  25. mosaic spread = 0.4º

  26. mosaic spread = 0.6º

  27. mosaic spread = 0.8º

  28. mosaic spread = 1.0º

  29. mosaic spread = 1.5º

  30. mosaic spread = 2.0º

  31. mosaic spread = 2.5º

  32. mosaic spread = 3.2º

  33. mosaic spread = 6.4º

  34. mosaic spread = 12.8º

  35. crystals bend!

  36. intensity  10 5 3 2.7 2.5 2.0 1.8 1.5 resolution (Å) stress and strain

  37. dispersion = 0

  38. dispersion = 0.014%

  39. dispersion = 0.25%

  40. dispersion = 0.6%

  41. dispersion = 1.3%

  42. dispersion = 2.6%

  43. dispersion = 5.1%

  44. intensity  10 5 3 2.7 2.5 2.0 1.8 1.5 resolution (Å) stress and strain

  45. Shutter Jitter open closed

  46. Shutter Jitter open closed

  47. Shutter Jitter open closed

  48. Shutter Jitter open closed

  49. Shutter Jitter open closed

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