Almost all protein crystallography groups have transitioned to robot mounting. The following instructions will demonstrate the complete workflow before, during, and after, your experiment for the average user.
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If you are new to the MX beamlines or your beamline induction has expired (check by logging into Portal, under Dashboard, Exams/Inductions) then you will need a beamline induction by a beamline scientist before starting your experiment.
Please e-mail beamline staff ahead of time to arrange a beamline induction. Typically, beamline inductions are run at 10am (MX1 experiments) and 2pm (MX2 experiments) at the start of your experiment. If you are cannot be present at these times, please let the beamline staff know ahead of schedule so that an alternative time can be arranged.
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In the MX Robot GUI (webpage), on the top right corner. If access is currently set to inactive, take over robot access by clicking on thebutton to change it to active.
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2. Add samples to your mount queue
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While the robot is mounting, the robot status will read:
'Goniometer motors locked for mount or collection'
“Prefetch” - robot prefetches your 2nd sample pin (sample second from the top) and places it in a slot that the robot can quickly retrieve.
While the robot is prefetching:
'Robot is busy but safe to collect'
It is safe to start collecting data whilst the robot is prefetching.
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If you need to go into the hutch. Make sure you ’Park' the robot. The robot status might read:
‘Robot is idle: Must be parked prior to hutch entry’
Avoid going into the hutch whilst the robot is moving, especially during mount and prefetch stages. For your safety, the robot will cease its movement when the hutch is unlocked and open and can cause it to stall. Parking the robot will take some time, when it is parked the following text should show up on the robot status:
‘Robot parked: Safe for hutch entry’
4. Sample Centering
At the top of the page, navigate to the ‘Sample Centering' tab. Most users prefer to use the 'Three Click Centering’ tool, just follow the prompts and pick a particular spot on your crystal.
The ‘Sample Position’ button allows you to move the centering by small amounts. Whilst the tweak angle button allows you to do small angle rotations via entering a custom step size.
5. Screening and Collecting
In the collection GUI, there are multiple tabs for data collection. You should use the ‘screening’ tab to screen your sample (20 degree wedge) to get an estimate on the resolution of your diffraction, the quality of the reflections, and possible space group and unit cell determination.
For larger crystals, it may be worth centering and screening multiple sites to find the best point for data collection
Once you are happy with your choice, navigate to the ‘collection’ tab. We recommend 1s exposure per 10 degrees (i.e. 36s exposure for 360 degree wedge). For most protein crystallography experiments, a 180 degree collection will be enough with proper sample centering and a good quality crystal, but when in doubt: collect 360 degrees.
The distance input determines the distance between your sample crystal and the detector. If your crystal diffracts all the way to the edge, consider moving the detector closer so that all reflections are accounted for. If the your crystal diffracts into a small area in the center, consider if your crystal is worth collection, and if so, move the detector further out.
6. Data Processing
The Data Processing page can be reached using any synchrotron browser by typing in:
processing/processing
It will show a list of auto-processed data by XDS for each of the samples that you have screened or collected. An excel spreadsheet export of all of the auto-process data is automatically uploaded to the EPN folder on our FTP server.
Finished collecting
When you have finished mounting and collecting your data, it is common courtesy and good beamline etiquette to 'Dismount and Park' the robot for the next user or for the beamline scientists.
It is good practise to communicate with your CAP group 15 minutes before you intend to finish to alert the next user that they can get ready to take over. If you are the last user for your experiment, it would be appreciated if you could reply to the beamline ready email and let the beamline scientists know that you are the last user and you are done with collection.