MX at Diamond Light Source
Katherine McAuley I03 PBS, Diamond
1 The aim of this talk is to give an introduction to synchrotrons and MX beamlines and how they work
2 Why do you need to collect MX data at synchrotrons?
Phasing experiments High resolu on High throughput Special Condi ons
3 What is a synchrotron?
A synchrotron is a par cle accelerator (typically electrons) which provides a source of extreme intensity light
Can be compared to a series of ‘super microscopes’ or a gigan c x‐ray machine;
Diamond is the UK’s synchrotron light facility that provides intense light ranging from Infrared to hard X‐rays
4 Synchrotrons in Europe
5 Synchrotron - main components
6 In this business, it's all about brightness !
X‐rays from Diamond are 100 billion mes brighter (1011) than from an X‐ray tube Or 10 billion mes brighter (1010) than the Sun >105 mes brighter than the previous UK Synchrotron, the SRS !
7 And Broad Continuous Spectrum!
8 Diamond is a 3rd generation light source
1st Genera on sources parasi c 2nd Genera on sources dedicated machines to synchrotron radia on emi ance typically high (150 nmrad) 3rd Genera on sources low beam emi ance use of inser on devices (wigglers and undulators) emi ance on order of 3‐20 nmrad
9 Bending Magnets curve the electron beam between adjacent straight sections and generate synchrotron light
Electron bunch
Synchrotron light
Number Wavelength (Å) Electrons of photons 10000 1000 100 10 1 0.1 1014
1013 Bending 12 10 magnet 1011
1010
109 X-ray tubes Emission spectrum 108 Sun 107 10 1 eV 10 eV 100 eV 1 keV 10 keV 100 keV Energy The MX beamlines at Diamond use insertion devices called undulators
Bending Bending magnet magnet 2 WigglerWiggler Electrons Brilliance (photons/s/mm 2/mrad2/0.1%BW) 3 UndulatorUndulator 1020
1019 3
1018
1017 2
1016
1015 1
1014 2 10 50 Energy (keV)
11 MX Village consists of 5 operational beamlines and several under development
Opera onal I02 I03 I04-1 Phase I I24 I04 . Tuneable I02, I03, I23 I04 Phase II . Fixed λ I04‐1 . Tuneable microfocus I24 In Development Phase III . Long λ I23 . VMX m and i
12 Beamlines consist of an optics hutch, experimental hutch and a control area
Booster synchrotron
LINAC
Storage ring
Beamline
13 The components in the optics hutch are used to select the X-ray energy and adjust the beam size
Storage ring
Op cs hutch
Experimental hutch 40 m
14 A double crystal monochromator is used to select the X-ray energy (wavelength)
Yaw Roll
nd 2 Crystal Beam Exit
‘Z’ Perpendicular Translation Vertical Pitch Offset
Beam ‘X’ In Parallel Translation θ Bragg Angle st ‘Y’ 1 Crystal Lateral Translation
Main Bragg Angle Rotation Figure 1 Axes Notation (‘Bounce Up’ Configuration Shown)
15 One mirror is used to focus the X-ray beam horizontally and another to focus vertically
16 The experiment and final beam conditioning happens in the experiment hutch
Storage ring
Op cs hutch
Experimental hutch 40 m
17 The end-station table supports:
CRLs A enuators Beam diagnos cs Slits On‐axis viewing system Sample environment
18 Beam conditioning
QBPM 16 Foil A enuators Diode – Shu er ‐ Diode
Compound Slits Refrac ve Lenses X Rays 19 The sample environment
Annealing device Backlight
Cryojet Backstop SPINE standard Viewing system pin Apertures
Fluorescence Detector Scatterguard
20 Fast pixel array detectors record the diffraction images
• Advantages • S/N much be er than CCD • Higher throughput • Fine phi slicing – be er data • Bea ng radia on damage – go even faster with high dose can collect more data at room temp • Key for CL3, protein dynamics, etc • On all beamlines since April 2013 • 100Hz on I03 and I24
21 At the end of each data collection, the data are automatically processed Data collection and experiment monitoring
Automated data reduction
Automated structure solution
22 Choose which beamline is appropriate to your experiment requirements Energy/wavelength? • Fixed energy – suitable? • Tuneable
Microfocus? • Use of apertures? • Dedicated microfocus beamline
Special Apparatus? • Dehydra on equipment • Spectroscopy • Biocontainment • In‐situ experiments
23 Use the web pages to compare the MX beamlines at Diamond
24 Beamline I02 I03 I04 I04‐1 I24 I23 VMXi VMXm Type Tuneable Op mised Tuneable μ‐ Tuneable Tuneable Tuneable μ‐ fixed focus long λ focus Beamsize [μm] 70 x 20 (mirrors) down 40 x 60 5 – 60 120 x 120 5 x 5 4 x 0.5 to 8 x 3 (CRLs) Flux [ph/s] > 1012 1011 > 1012 > 1012 > 1012 (DCM) > 1013 > 1014 (DMM) Pilatus Detector P6M P3‐6M P6M P2M P3‐6M Custom TBD TBD (25 Hz) (100 Hz) (25 Hz) (30 Hz) (100 Hz) P12M Sample changer Rigaku ACTOR (< 35 s) Irelec CATS (< 45 s) Custom TBD TBD made Containment CL 1 CL 3 CL 1 CL 1 CL 2 CL 2 CL 2 CL 2 Humidity On request No TBD No Controller Mul axis Mini‐ Mini‐kappa Custom No No goniometry kappa kappa Minibeam Yes Development apertures In situ Soon Yes Yes Yes No Yes TBD Crystal Washer Yes Yes Yes Yes Soon No No No and Annealer Remote Access Yes No Yes TBD
25 Techniques available on the MX beamlines Using fluorescence spectra to identify metals in your sample
All beamlines have fluorescence detectors Take a fluorescence spectrum of your sample in seconds Automa c iden fica on of your sample with summary in webpages:
27 If there is a metal present:
On the tuneable beamlines (i.e. not I04‐1) collect fluorescence scan around the edge e.g. Signal from Br soaked lysozyme Results stored in webpages/ ISPyB:
28 The Mini Kappa can be used to reorientate crystals for data collection
Before reorienta on
A er reorienta on Op mised MAD data collec on; Bijvoet pairs on the same frame Improved high‐mul plicity SAD data collec on protocols Smart data collec on strategies, be er completeness of data, especially P1 Reducing/avoiding spot overlap, be er spot separa on (long unit cell axes) Mul ‐crystal data collec on: collect missing data Help in point group determina on Comparing crystals in the same orienta on
29 The HC1 device controls sample humidity and can improve crystal diffraction
30 In-situ crystallography: data collection from crystals in crystallisation plates
I03
I24
I04‐1
31 Pathogenic samples can be studied on I03 and I24
All beamlines level 1 I03 and I24 up to level 2 biological containment Cryo and in‐situ samples I03 containment level 3 compliant In‐situ samples only Request via standard beam me applica on process
32 Coming soon to the MX Village (new developments)
33 Performance: Crystal preparation 500 crystals in 2-day experiment
Recent 34 Prepara on Collec on (Full analysis: 2‐7 days) experiment: 200 crystals in 6 hours Overnight Unattended data collection queues Sample auto‐ centring: >97%
10am midnight 9am
Typical
Experienced
Una ended >350 crystals
35 Coming soon to the MX Village (new beamlines)
36 I23 - Long wavelength Macromolecular Crystallography Beamline
I23 will be the first MX beamline op mized for the long‐wavelength region (1.5 – 4 Å).
It will provide a unique tool to fully exploit the poten al of experimental phasing from na ve protein and DNA/RNA crystals.
Sample changer
Detector readout electronics box
Omega axis
Diffraction detector
37 VMXmicron Sub micron variable focus
Aims Challenges
0.5 x 0.5 um – 5 x 5 um sample delivery beam sample visualiza on variable aspect ra o DCM rapid beam size change 13nrad pitch stability 5 – 30 keV (dependent on ID choice) op cs fast sample exchange/ facilita ng fast beamsize delivery changes is the real challenge fast crystal loca on 2014 Team • Technical Design Report • Gwyndaf Evans
• 38 Prototyping • Jose Trincao VMXi - dedicated to in-situ (plate) crystallography This beamline will provide Plate storage facility Automated transfer between storage and beamline Advanced laser imaging of drops (on‐line and off‐line) X‐ray data collec on and screening (automated or interac ve) Tuneable microfocus beam (5x5 micron – 10‐25 keV) Narrow or broad bandpass beam (2eV – 100eV) Diffrac on data available less than 48 hours a er shipping plate to Diamond ‐ during run me…
39 MX Village Support Team
I02 I03 I04 I04‐1 I23 I24
Thomas Robin Owen Sorensen Katherine Frank von Del McAuley Dave Hall Armin Wagner Juan Sanchez‐ Danny Axford Weatherby Jose Brandao Darren Sherrell Vitaliy James Sandy Stuart Fisher Ralf Flaig Mykhaylyk Alice Pierre Aller Douangamath Carina Lobley Anna Warren Pierpaolo Mark Williams Ramona Duman Marco Romano Petra Lukacik Mazzorana Neil Pa erson
40 MX Village Support Team
Data Data Industrial MX Controls Engineering EHCs Acquisi on Analysis Team Technicians
Ted Cassidy Dave Butler Mar n Burt Jon Blakes Alun Ashton Ronaldo Mercado Elizabeth Sho on Les Clinker Mic Harding Alistair Donaldson Graham Duller Nick Gorringe Thomas Hartrampf Paul Hathaway Karl Levik James O’Hea Alex Dias Doug Sco Adam Presco Mar n Gilbert Nathan Sear Adam Taylor Paul Symes Chris Sharpe Graeme Winter Andy Foster Jitka Waterman Geoff Preece Russell Walker Tim Whitewood
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