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Continuous tilt electron diffraction recording » History » Version 24

Anchi Cheng, 10/28/2020 07:30 PM

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h1. Continuous tilt electron diffraction recording use Ceta or Ceta-D camera and TFS scopes
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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.
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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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h2. Components of the feature
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# Diffraction mode TEM instrument
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# Beamstop control: achieved by AutoIt Scripts BeamstopIn and BeamstopOut
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# Rolling-shutter movie acquisition using TIA interface
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# [[TIA raw data conversion|Data conversion]] and upload of the movies into Leginon database and crystallography format
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# [[ MSI-Diffr application ]]
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# [[Camera length calibration]]
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h2. Installation (assuming that you've already have Leginon installation).
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# Check that you have access to Advanced TEM Scripting (frame-saving upgrade not needed).
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# 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 and webserver for myamiweb.
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# import updated Calibrations application (see [[Steps_involved_in_the_installation]])
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# 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)
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# [[Setup Diffraction mode TEM instrument]]
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# [[Setup Ceta to use Advanced TEM Scripting]]
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# [[AutoIt program and script compilation]]
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# [[Setup movie upload with diffrtransfer.py]]
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h2. Usage
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h3.  Check these before you start:
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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.
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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]]. 
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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]]
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h3. Start MSI-DIffr application and assign client as required.
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h3. [[Setup df preset]]
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h3. Grid atlas collection
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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.
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2. Select a larger C2 aperture for grid atlas collection.
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3. collect grid atlas.
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4. Reselect the small C2 aperture.  Make sure it is still centered.
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h3. Queue up potential crystals
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1. Pick square and then pick "hole" for intermediate mag image of potential crystals.  (This is no different from any other MSI application).
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2. Submit preview targets in "DExposure Targeting" node for a small rotation image and confirm whether it diffracts or not.
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3. Submit the good crystal positions as acquisition targets into the queue in "DExposure Targeting" node.  No focus target is needed.
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4. If needed, modify "DExposure" settings for start, range, and speed of the tilt.
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5. Click "process queue" tool to start collection
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h3. Diffraction series naming convension
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The diffraction series is named after the parent image database id and target number in its target list.  For example, 123456_1