Continuous tilt electron diffraction recording » History » Version 20
  Anchi Cheng, 04/29/2020 07:04 PM 
  
| 1 | 6 | Anchi Cheng | h1. Continuous tilt electron diffraction recording use Ceta or Ceta-D camera and TFS scopes | 
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| 2 | 1 | Anchi Cheng | |
| 3 | Micro-ED involves recording 3D crystal electron diffraction pattern during a continuous stage tilt. Leginon implementation of this application includes the listed components below and is available by using myami-ed branch until it is officially added to future release. As is, it is only fully implemented for TFS Ceta or Ceta-D camera. | ||
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| 5 | Since many of the feature required here are not available through Standard or Advanced TEM Scripting from TFS, AutoIt scripts are used in several places. | ||
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| 7 | h2. Components of the feature | ||
| 8 | |||
| 9 | 5 | Anchi Cheng | # Diffraction mode TEM instrument | 
| 10 | 8 | Anchi Cheng | # Beamstop control: achieved by AutoIt Scripts BeamstopIn and BeamstopOut | 
| 11 | 9 | Anchi Cheng | # Rolling-shutter movie acquisition using TIA interface | 
| 12 | # Data conversion and upload of the movies into Leginon database and crystallography format | ||
| 13 | 4 | Anchi Cheng | # [[ MSI-Diffr application ]] | 
| 14 | 11 | Anchi Cheng | # [[Camera length calibration]] | 
| 15 | 1 | Anchi Cheng | |
| 16 | h2. Installation (assuming that you've already have Leginon installation). | ||
| 17 | |||
| 18 | # Check that you have access to Advanced TEM Scripting (frame-saving upgrade not needed). | ||
| 19 | 12 | Anchi Cheng | # Install or set environment to use git branch myami-ed or current myami-beta on TFS microscope with Ceta camera as well as your leginon linux box. | 
| 20 | 10 | Anchi Cheng | # import updated Calibrations application (see [[Steps_involved_in_the_installation]]) | 
| 21 | # import MSI-diffraction and settings from your_myami/leginon/applications (see general description of [[Steps_involved_in_the_installation]] regarding importing application and additional settings for the application) | ||
| 22 | 5 | Anchi Cheng | # [[Setup Diffraction mode TEM instrument]] | 
| 23 | 7 | Anchi Cheng | # [[Setup Ceta to use Advanced TEM Scripting]] | 
| 24 | 3 | Anchi Cheng | # [[AutoIt program and script compilation]] | 
| 25 | 20 | Anchi Cheng | # [[Setup movie upload with diffrtransfer.py]] | 
| 26 | 2 | Anchi Cheng | h2. Usage | 
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| 28 | 15 | Anchi Cheng | h3. Check these before you start: | 
| 29 | 1 | Anchi Cheng | |
| 30 | 15 | Anchi Cheng | 1. Confirm that instruments.cfg is properly set up to use feicam for Ceta camera. Especially if you use Falcon camera on the same instrument for imaging. | 
| 31 | 18 | Anchi Cheng | 2. Open TUI. Select and insert BM-Ceta in the camera tab. Confirm that tui_acquire.au3 validation items are set [[Autoit program and script compilation#TUIAcquire-Testing|correctly]]. | 
| 32 | 19 | Anchi Cheng | 3. Make sure TIA window is available and the Export Series shortcut is shown as required by [[Autoit program and script compilation#TiaExportSeries-Testing|TiaExportSeries.au3]] | 
| 33 | 15 | Anchi Cheng | |
| 34 | 13 | Anchi Cheng | h3. Start MSI-DIffr application and assign client as required. | 
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| 36 | 1 | Anchi Cheng | h3. [[Setup df preset]] | 
| 37 | 15 | Anchi Cheng | |
| 38 | h3. Grid atlas collection | ||
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| 40 | 1. Select gr preset to scope, with flucam showing the grid and aperture, draw on the computer screen the location of the centerred small C2 aperture. | ||
| 41 | 2. Select a larger C2 aperture for grid atlas collection. | ||
| 42 | 3. collect grid atlas. | ||
| 43 | 4. Reselect the small C2 aperture. Make sure it is still centered. | ||
| 44 | |||
| 45 | h3. Queue up potential crystals | ||
| 46 | |||
| 47 | 1. Pick square and then pick "hole" for intermediate mag image of potential crystals. (This is no different from any other MSI application). | ||
| 48 | 2. Submit preview targets in "DExposure Targeting" node for a small rotation image and confirm whether it diffracts or not. | ||
| 49 | 3. Submit the good crystal positions as acquisition targets into the queue in "DExposure Targeting" node. No focus target is needed. | ||
| 50 | 4. If needed, modify "DExposure" settings for start, range, and speed of the tilt. | ||
| 51 | 5. Click "process queue" tool to start collection |