If you are using Krios
LOCATION: Diamond Light Source
Harwell Science and Innovation Campus
Krios I: Currently in the synchrotron building between beamlines I20 and I19.
From Diamond House, enter the ring, turn left and down the first set of stairs. Turn left as you exit the stairwell for a few meters and Krios I will be on your right.
Krios II: Currently in the synchrotron building between beamlines I13 and I14.
From Diamond House, enter the ring, turn right, and continue to the Zone 10 stairwell. Go down the stairs and turn left as you exit the stairwell. You will need to cross over the I13 beam pipe via the metal stairs, and Krios II will be on your right.
If you need assistance please contact the Experimental Hall Coordinators on 0044 (0)1235 778787.
Directions to Diamond itself can be found here.
If you are using the Polara
See OPIC website for more information.
Successful Rapid proposals will be scheduled time on the microscope which will be allocated in 48 hour slots. However for Rapid sessions scheduled on a Friday 72 hours may be given depending on microscope cryo-cycles. Users will be expected to be onsite during the office hours of the first day and provide enough samples for automated data collection to run for the following 40 hours.
For BAG proposals sessions may be longer than 48 hours. However we recommend that each sample is given a 48 hour slot. Preferably all samples will be loaded on the first day of the BAG session but new samples can be loaded every 48 hours.
Data Storage: Users are requested to bring 3x2TB hard drives.
Samples: All proposals require evidence of suitable cryo EM samples (see Acceptance Criteria). Currently we do not have the facilities to offer onsite sample preparation and request that users bring sufficient grids for their full allocation of instrument time.
Principal Investigators will be prompted to submit the experimental team and sample information via the User Administration System (UAS). These will be processed and the PI will receive the details.
Please remember to put all samples you are bringing to Diamond on the ERA, as samples not on the ERA will NOT be loaded into the microscopes.
At the microscope
Users of both Krios 1 (m02) or Krios 2 (m03) should meet their local contact at Krios 1 as grid loading is carried out for both microscopes in lab. 7 of zone 1. The standard user day is listed below:
- 9 - 9.30: Session starts
- 9.15 - 10.15: Load up to 8 grids into auto grid cartridges and the Krios cassette
- 10.20 - 10-30: Load auto grid cassette into the Krios
- 10.30 – 12: Survey grids to determine the best grid
- 12 – 12.45: Collect grid atlas image
- 12.45 – 3: Setup EPU/TOMO4, microscope alignments, detector gain references
- 3 – 4: Check images, assess CTF and particle density.
- Automatic data collection for ~40 hours.
EBIC staff will check data acquisition is proceeding from home but the user should also monitor their data collection and email their local contact if they observe issues.
If you have written your data to USB HDDs please make sure that your local contact has the correct mailing address for return postage. Alternatively FTP/Globus may be used to transfer your data from Diamond to your home institution.
Users publishing work containing data from eBIC should ensure they:
- Acknowledge DLS and any of its personnel in any published material that results from their use of the Cryo-EM facilities. The following statement should always be included in any publications:
“(We acknowledge) Diamond for access and support of the Cryo-EM facilities at the UK national electron bio-imaging centre (eBIC), proposal EM####, funded by the Wellcome Trust, MRC and BBSRC.”
- Notify DLS of the title, authors, facility, journal and reference number, date of acceptance and publication of any scientific publications or papers as a result of their use of the beamtime
- Notify DLS if they plan to publish any materials e.g. general reports, press releases, etc. that result from their use of the beamtime prior to the material being published and allow time for DLS to review such material and give its prior permission to publish such material if reasonably requested by DLS.