waretomo 0.1.0

Overengineered batch processing script for tomography data with Warp and aretomo.

waretomo

Overengineered batch processing script for tomography data with Warp and aretomo.

Installation and usage

pip install waretomo

waretomo -h

Walkthrough

Here's a short summary of how I recommend using this script:

• pre-process your dataset in Warp, up to Create stack for imod. Make sure to deselect bad tilts from the warp interface; they will be skipped in subsequent steps
• run waretomo in dry-run mode (-d), to make sure everything is set up correctly. Pass any other options you'd like (see waretomo -h for a complete list and shorthands):

waretomo . --mdoc-dir ./mdocs  -b 8 -t 800 -d

• follow error messages if any arise (you may have to provide the path to the AreTomo executable with --aretomo, for example)
• once the above command works, it will give you a summary of inputs and of the upcoming pipeline. It's time to try to run it on a single tomogram, using -j (--just). Note that the name to provide to -j is the tomogram name given at the top of the mdoc file (ImageFile = <something>), and not the input file (this is because warp actually uses this value for all subsequent outputs).

waretomo . --mdoc-dir ./mdocs  -b 8 -t 800 -j tiltseries_23.mrc

• this will run the full pipeline on that tomogram. Keep an eye out for error messages, they might give you tips for solving them.
• if everything works, you will now have a few outputs to check out:
  • ./waretomo_processing/tiltseries_23.mrc: raw aretomo reconstruction.
  • ./waretomo_processing/denoised/tiltseries_23.mrc: same as above, denoised with topaz for better annotation
  • <mdoc-dir>/mdoc_tilted/tiltseries_23.mrc.mdoc: mdoc file updated with skipped tilt and with adjusted tilt angles from aretomo's TiltAlign option (e.g: to align lamellae to the XY plane)
  • ./waretomo_processing/tiltseries_23.xf: alignment metadata, used by warp together with above mdocs for reconstruction.
• check out the reconstructions and make sure everything looks as you want. If anything is wrong, adjust parameters as you see fit. You can also run only parts of the script by using the --start-from and --stop-at options (both are inclusive). Use -f if you want to overwrite existing outputs.
• once you're happy, remove the -j option to process the full dataset.

At this point, you're ready to go back to Warp. Here, you can simply import tilt series from IMOD. Don't forget to provide the mdoc_tilted directory instead of the original mdocs. Set waretomo_processing as the Root folder with IMOD processing results, and Warp will find the xf files located there. Provide the pixel size of the binned aretomo reconstructions (find out with e.g: header waretomo_processing/tiltseries_23.mrc.mrc). You might have to provide the dose per tilt, depending on the origin/correctness of your mdocs.

You can now proceed with reconstructing with Warp, use M, and so on, while having matching AreTomo reconstructions with their local patch alignments and subsequent denoising to maximize readability for picking and annotation.