Microfocusing Xray Protein Crystallography Beamline Zengqiang Gao beamline

Microfocusing X-ray Protein Crystallography Beamline Zengqiang Gao beamline 17/12/2019 The 2 nd Meeting of HEPS International Advisory Committee High Energy Photon Source - Beijing

Table of Contents • Scope and objective • Outcome of the review 2018 • Status of work progress • Summary of authorized changes • Manpower • Risks and mitigation • Schedule and major upcoming milestones HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 2

Scope and objective ØDetermine the crystal structures of vital proteins in life sciences S. F. , Yang et al, Nature, 560, 666 -670 l Membrane proteins (GPCR et al) l Huge complex (ribosome et al) l Other proteins Ferbitz, L. et al, Nature, 431, 590 -596 ØServing as the platform for drug discovery ØHigh throughput/automatic HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 Hubbard, R. E. , J. Synchrotron Rad. 15, 227– 230 3

Table of Contents • Scope and objective • Outcome of the review 2018 • Status of work progress • Summary of authorized changes • Manpower • Risks and mitigation • Schedule and major upcoming milestones HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 4

Outcome of the review 2018 Type Comment Description Response Protein crystallography is a continuously developing field. Look carefully around the world status and choose best-possible endstation system in the latter half of the project. In contrast, the beamline is more or less standard which can be finalized in an earlier stage All the components of MX beamline will be installed at the end of 2023, and the commissioning with x-ray beam will be start at the beginning of 2024 The heat-load level is manageable with cryogenic silicon monochromator without the white beam mirror. Less numbers of optical components can make the beam stability better and make the life easier. If it is not enough, consider a diamond window instead of the WBM White beam mirror has been removed The beamline optics goal may be reached by the combination of only the HDCM and CRLs. Since the CRL is a chromatic component, design a mechanism to fix the focal spot. The mechanism may also be used for changing the beam size by defocusing. We use HDCM and K-B mirror to get the 1µm spot and keep higher flux at the same time. CRL is used to get the larger spot (30µm) HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 5

Outcome of the review 2018 Type Comment Description Response Most likely, the combination of cryo-sample with serial Room temperature/in situ with crystallography is not necessary. Room temperature measurement does SSX will be the dominating not necessarily couple with the low range of degree especially for serial experimental method. crystallography From the users’ point of view, it is comfortable to use the same software (preferably the software pipeline) between SSRF and HEPS. The ideal situation may be to set up a consortium to develop the software pipeline among SSRF, HEPS, TPS, PLS and SPring-8 The data processing pipeline will be same as, even identical with, that of SSRF. The data collection software (GUI) maybe different from that of SSRF. Serial crystallography for LCP based membrane proteins may not be fit Fixed-target system or Highwith ‘jet’-scheme. Consider a new method for the LCP based viscosity extrusion (HVE) crystallography injector will be used for LCP based proteins. HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 6

Table of Contents • Scope and objective • Outcome of the review 2018 • Status of work progress • Summary of authorized changes • Manpower • Risks and mitigation • Schedule and major upcoming milestones HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 7

Status of work progress — optical layout Energy resolution: 1. 6/12400=1. 3× 10 -4 Spot size(FWHM, H*V) 41. 2µm*17. 6µm Shielding wall Spot size(FWHM, H*V) 0. 7µm*0. 8µm Mono slit CRL White slit VFM HFM sample l Energy range: 5 -18 ke. V l Energy resolution(∆E/E): ≤ 2× 10 -4@12. 4 ke. V l Spot size(FWHM, µm): 1 -30 (discrete) l Flux : ~7 1013 phs/s@12. 4 ke. V undulator HDCM 31. 5 m 43 m 45 m 49. 5 m 50 m 52. 56 m 52. 9 m Design principle: as less optics as possible 1µm spot: HDCM + KB mirror Larger spot: HDCM + KB mirror + CRL 53. 36 m Spot divergence(FWHM, H*V) 1. 2 mrad*0. 8 mrad Spot divergence(FWHM, H*V) 11. 6µrad*12. 6µrad HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 8

Status of work progress — Light source Storage ring energy 6 Ge. V current emittanc e x y Energy spread 200 m. A 60 pm. rad Type Total length IAU undulator Phase error（ RMS） 5 m Brightness (@12. 4 ke. V) 6. 5 1021 phs/s·mrad 2·mm 2· 0. 1%BW 2. 1 1015 phs/s· 0. 1%BW 14. 3 k. W 10. 12 m Periodic length 32. 7 mm flux (@12. 4 ke. V) 9. 64 m 1 st order energy 2. 647. 9 ke. V Total power 1. 06 E-3 K value 0. 69 -2. 43 x/ x 18. 5 m/1. magnetic 0. 8 T 83 rad field y/ y 6. 2 m/ 0. 64 rad Min gap 11 mm Center power density Beam size（ FWHM， @12. 4 ke. V） Beam divergence（ FWHM， @12. 4 ke. V） HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 4° 336 k. W/mrad 2 1 st 3 rd 5 th Flux curve Brill. curve 41. 1μm (H) × 17. 4μm (V) 13. 3μrad (H) × 12. 8μrad (V) Power density distribution 9

Status of work progress — HDCM Parameter (unit) Distance to light source (m) Accept divergence (μrad) Crystal surface Energy (ke. V) Bragg angle ( ) Designed bragg angle ( ) type Reflection direction Offset (horizontal, mm) Offset deviation(µm) Max slope error due to heat (RMS, μrad) Cooling method Base material Value 45 22× 22 Si<111> 5~18 6. 31~23. 29 -4~30 Fixed offset horizontal 20 ∆E/E = 1. 6/12400=1. 29× 10 -4 (Xrt) ∆E/E = 1. 25× 10 -4 (XOP) Max temperature: 147 K@2. 6 ke. V Energy resolution: ~1. 3× 10 -4 Shape/slope error deformation (meridian)： 0. 06 nm/0. 32 rad Shape/slope error deformation (sagittal)： 0. 018 nm/0. 35 rad ≤ 10 0. 32 LN 2 granite Next plan: replace the DCM with channel-cut monochromator (CCM), theoretical calculation is going on. HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 10

Status of work progress — KB mirror parameter（unit） Object distance（m） Accept divergence (μrad) type Image distance（m） Energy range（ke. V） Mirror size(L×W×H， mm 3) Sagittal slope error（ rad） Meridian slope error（ rad） roughness（Å） Surface direction Meridian type Reflection angle（ mrad） coating VFM 52. 56 17× 17 HFM 52. 9 17× 17 Fixed shape 0. 8 0. 46 5～ 18 300× 50× 50 1 1 0. 2 3 Horizontal Cylinderial 3 3 vertical Cylinderial 3 Rh Rh HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 KB mirror is also used for high order harmonic rejection: Si (the mirror surface) for 1 st order Rh for 3 rd and 5 th order Spot size: 0. 68 m(H)× 0. 76 m(V); Flux: 7. 11× 1013 ph/s Tracing result with DCM shape error by heat and KB slope error 11

Status of work progress — CRL Parameter (unit) Object instance (m) Accept divergence (μrad) Radius at apex ( m) Geometry radius ( m) Material Number of lens Value 50 17× 17 200 900 Be 6 Two methods to change spot size： Ø Change the number of CRL lens Ø Use all lenses, change the size by pinhole R=200，N=1 R=200，N=3 N of lens 1 2 3 4 5 6 Spot size (H×V, m) 3. 2× 9. 0 7. 0× 18. 4 10. 0× 29. 0 14. 8× 32. 4 22. 0× 42. 8 25× 48. 2 Flux (ph/s) 6. 78× 1013 6. 58× 1013 6. 41× 1013 6. 22× 1013 6. 05 × 1013 5. 89 × 1013 R=200，N=6 The beam distribution becomes more uniform with the number of lens increase HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 12

Slope error and stability analysis Angular stability Optical elements parameters: Position stability Depends on the Magnification factor (p/q) Optical element HDCM VFM HFM Deviation ratio (H/V) Slope error (M/S) 0. 3μrad/1μrad 0. 2μrad/1μrad Angle deviation 50 nrad 80 nrad 1. 3%/ 3. 1%/ 2. 2% 5. 2% Angle deviation 100 nrad 4. 8%/ 8. 0% Angle deviation 150 nrad 110 nrad 150 nrad 10. 2%/ 9. 9% HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 LDCM=45 m, LVFM=52. 56 m, LHFM=52. 9 m, MVFM=65. 7, MHFM=115 name H/meridian Light source HDCM angle deviation VFM position deviation HFM angle deviation HFM position deviation beam size Without deviation beam size With deviation Deviation ratio (rms) 41. 1μm Deviation at Sample 0. 036μm 17. 4μm Deviation at Sample 0. 026μm 0. 1μrad 0. 18μm 0. 1μrad 0. 08μm 0. 1μrad 0. 57μm 3μrad 0. 38μm - - 1μm 0. 02μm 0. 1μrad 0. 22μm 3μrad 0. 03μm 1μm 0. 01μm - - V/sagittal 0. 93μm 0. 95μm 0. 97μm 1. 03μm 4. 8% 8. 0% 13

Status of work progress — Experimental methods Conventional crystallography Common (low temperature， loop) In-situ (room temperature，plate) Serial Fixed-target sample delivery crystallography Jet-based sample delivery Local access HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 First stage Key area of development Remote access 14

Status of work progress — endstation hardware Yaw KB Mirror Z Y Roll Detector X X-ray in detector： Frame rate：>100 Hz diffractometer Pitch X-ray out Eiger Robot Detector developed by IHEP diffractometer: MD 3 -up Sphere of confusion (SOC): 0. 1μm Layout of experimental endstation (by Dr. Hao, Liang) Generals hardwares will be available through the commercial products. HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 Robot: ISARA Time for sample change: ≤ 10 s Success ratio: ≥ 99. 99% 15

Hardware-Sample delivery system S. Sui et al. , Struct Dynam. Us 4, 2017, 032202 Jet-based An employee will be hired to develop this kind of instrument in next year! Fixed target G. R. Guo et al, IUCr. J (2018). 5, 238– 246 Only be used as a holder C. Y. Huang et al, Acta. D, 2015, 1238 In-situ, fewer crystals P. Roedig et al. , nature methods, 2017, 14, 805 In-situ, many crystals Fixed target system (Chip-based) will be the first choice for SSX experiment in HEPS (at least in the first stage). HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 16

Status of work progress — software http: //mxcube. github. io/mxcube/ Integrated new hardware control into the original MXCu. BE GUI successfully! The data collection software (GUI) has been identified to be MXCu. BE HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 17

Status of work progress — software Data processing software l Data process: HKL 2000/3000, Xia 2(ccp 4); XDS; Fast_dp (Diamond); Go. com (SLS); l Phase determination: Phenix. autosol; Phaser; Mr. BUMP: Auto. SHARP l Serial data process: XDS/Crystal. FEL/facility unique K. Yamashita et al, Acta Cryst. D, 2018, 74, 441– 449 individual pipeline F. Yu et al J. Appl. Cryst. (2019). 52, 472– 47 Standard data: using the current software (integrate it into pipeline) SSX data: develop our own software based on current one such as Cryst. FEL HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 18

Table of Contents • Scope and objective • Outcome of the review 2018 • Status of work progress • Summary of authorized changes • Manpower • Risks and mitigation • Schedule and major upcoming milestones HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 19

Summary of authorized changes wall IAU WBM CRL Pre-focus K-B Focus K-B aperture sample SSS 2018 Optimization of optical layout Ø From two-step focusing to focus directly Ø Less number of optical elements (6/7 to 3/4) Ø Shorter whole length of beamline (78. 7 m to 53. 4 m) Shielding wall undulator Mono slit White slit Ø Control software (GUI) has been identified CRL VFM HFM sample HDCM 31. 5 m 43 m 45 m (MXCu. BE) 2019 49. 5 m 50 m 52. 56 m 52. 9 m 53. 36 m HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 20

Table of Contents • Scope and objective • Outcome of the review 2018 • Status of work progress • Summary of authorized changes • Manpower • Risks and mitigation • Schedule and major upcoming milestones HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 21

Manpower HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 22

Table of Contents • Scope and objective • Outcome of the review 2018 • Status of work progress • Summary of authorized changes • Manpower • Risks and mitigation • Schedule and major upcoming milestones HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 23

Risks and mitigation Risk ID Description the time for getting high BA-RI-01 quality KB mirror maybe longer than expected monochromator will be BA-RI-02 self-developed, schedule maybe tight BA-RI-03 quality of CRL lens can not be controlled effectively Date Identified Likeliho od Risk Class (severity) Control Measure Risk Owner Status Date Closed 2019/01 Medium estimating the potential vender, such as Zeiss et al. BA/Optical system Ongoing Not sure 2019/01 Medium more staffs BA/Optical system Ongoing Not sure 2019/01 Low test in advance, try to choose the product with reliable quality BA/Optical system Ongoing Not sure HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 24

Table of Contents • Scope and objective • Outcome of the review 2018 • Status of work progress • Summary of authorized changes • Manpower • Risks and mitigation • Schedule and major upcoming milestones HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 25

Schedule and major coming milestone Major coming milestone Ø 2020. 06. 30 the optical layout design frozen Ø 2020. 12. 31 the engineering design frozen Ø 2022. 03. 31 start to Experiment Hall alignment Ø 2023. 01. 02 start to install and commissioning the beamline equipment offline Ø 2024. 01. 02 start to beamline commissioning with X-ray beam Class I CPM plan of MX beamline HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 26

Summary Ø Ø Ø Optimize the optical layout based on the recommendation of 2018 IAC committee Ø From two-step focusing to focus directly Ø Will replace the HDCM with HCCM Identify the control software (MXCu. BE) The experimental method will focus on serial synchrotron crystallography (SSX) HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 27

Acknowledgement l The support team (optical, mechanics, control, engineering drawing etc) l SLS: Prof. Meitian Wang SSRF: Prof. Jianhua He, Dr. Qisheng Wang, Dr. Bo Sun l HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 28

Thank you for your attention! HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 29

Supporting material Compare HEPS-MX beamline to others micro-focusing beamlines HEPS-MX Energy range （ke. V） Spot size Energy resolution flux (phs/s@1× 1μ m) status Micro. MAX（ MAXIV） VMXm (Diamond) BL 02 U (SSRF) BL 32 XU (Spring-8) ID 29/EBSL 8 (ESRF) 5 -18 5 -30 7 -25 7 -15 9 -15 10 -30 1 -10 0. 5 -2 1 -15 0. 5 -10 7 e 10 2 e 14 (0. 3%BW) 2 e-4 7. 0 e 13 Under design 2 e-4 1. 0 e 13 Under design HEPS · The 2 nd Meeting of HEPS IAC, IHEP, Dec. 16 -18, 2019 ~1. 0 e 12 In operation 3. 5 e 11 Under design In operation Under design 30