Continuous tilt electron diffraction recording » History » Version 26
  Anchi Cheng, 11/05/2020 08:02 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 | 22 | Anchi Cheng | 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-beta branch until it is officially added to future release. As is, it is only fully implemented for TFS Ceta or Ceta-D camera. | 
| 4 | 1 | Anchi Cheng | |
| 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 | 23 | Anchi Cheng | # [[TIA raw data conversion|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 | 25 | Anchi Cheng | # Install or set environment to use git branch myami-3.5 ant up or the current myami-beta on TFS microscope with Ceta camera as well as your leginon linux box and webserver for myamiweb. | 
| 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 | 21 | Anchi Cheng | |
| 27 | 26 | Anchi Cheng | h2. Presets unique for this application | 
| 28 | |preset name|TEM|magnification(camera length)|SpotSize| | ||
| 29 | |hl|Glacios|8500|10| | ||
| 30 | |df|DiffrGlacios|1100|10| | ||
| 31 | |||
| 32 | h2. Calibration | ||
| 33 | # [[Camera length calibration]] | ||
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| 35 | 2 | Anchi Cheng | h2. Usage | 
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| 37 | 15 | Anchi Cheng | h3. Check these before you start: | 
| 38 | 1 | Anchi Cheng | |
| 39 | 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. | 
| 40 | 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]]. | 
| 41 | 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]] | 
| 42 | 15 | Anchi Cheng | |
| 43 | 13 | Anchi Cheng | h3. Start MSI-DIffr application and assign client as required. | 
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| 45 | 1 | Anchi Cheng | h3. [[Setup df preset]] | 
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| 47 | h3. Grid atlas collection | ||
| 48 | |||
| 49 | 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. | ||
| 50 | 2. Select a larger C2 aperture for grid atlas collection. | ||
| 51 | 3. collect grid atlas. | ||
| 52 | 4. Reselect the small C2 aperture. Make sure it is still centered. | ||
| 53 | |||
| 54 | h3. Queue up potential crystals | ||
| 55 | |||
| 56 | 1. Pick square and then pick "hole" for intermediate mag image of potential crystals. (This is no different from any other MSI application). | ||
| 57 | 2. Submit preview targets in "DExposure Targeting" node for a small rotation image and confirm whether it diffracts or not. | ||
| 58 | 3. Submit the good crystal positions as acquisition targets into the queue in "DExposure Targeting" node. No focus target is needed. | ||
| 59 | 4. If needed, modify "DExposure" settings for start, range, and speed of the tilt. | ||
| 60 | 5. Click "process queue" tool to start collection | ||
| 61 | 24 | Anchi Cheng | |
| 62 | h3. Diffraction series naming convension | ||
| 63 | |||
| 64 | The diffraction series is named after the parent image database id and target number in its target list. For example, 123456_1 |