Prepare gridsquare (set eucentric height)Edit Delete
|Name||Prepare gridsquare (set eucentric height)|
|Category||Other Data Acquisition|
|Description||Operating on points saved in the Navigator, this script automatically sets the eucentric height of each gridsquare by using the Rough Eucentricity routine in SerialEM at two different magnifications.|
|Author||Jason de la Cruz|
|Upload date||June 30, 2017|
|SerialEM version ≥||3.7|
Modify settings in section below. Set "setIntensityCmd" to either "SetIlluminatedArea" for FEI Titan series or "SetPercentC2" for all other microscopes. Decide on condenser beam illumination (aperture, spot size); enter desired spot size in the settings section. Then set "highMag" to a mag that maximizes area of single gridsquare on detector. Adjust intensity for optimum signal, center beam shift, then record intensity value in "highMagIntensity".
Then drop the mag to lowest mag with unobstructed view of TEM grid (i.e., the mag you use for whole-grid atlas), center beam shift, adjust intensity, and enter appropriate values in "lowMag" and "lowMagIntensity".
Open Navigator and collect whole-grid atlas. Add a point at the center of each gridsquare you wish to set at eucentric height.
If necessary, go to each XY coordinate to center the point in "highMag" using the Navigator "Move Item" button. For those points you want to run the script on, set to "Acquire".
When ready to start, open file to save autodoc metadata (with .mdoc or .idoc). Run the script using the SerialEM menu option "Navigator > Acquire at Points". Close the file when done.
Please check that beam is centered in both low and high mags before continuing.
Ensure all checkboxes in "Script > Controls..." dialog are unchecked to avoid "low counts" errors.
Copyright (c) 2017, Howard Hughes Medical Institute. License ISC: Internet Software Consortium license, http://opensource.org/licenses/ISC. This is free software: you are free to change and redistribute it. There is NO WARRANTY, to the extent permitted by law.