Reference based motion correction with micrographs from two different microscopes

Data Processing Strategy
1

Hello everyone,

Apologies if this has been posted before but I couldn’t find it. I have particles that come from two different microscopes for my project, and I would like to use reference based motion correction to improve the final map. Typically what I do use MotionCorr2 to bin my micrographs exactly by the amount they need to match the particle size of my other movies and import those into Cryosparc for data processing. However to use reference based motion correction I would need to have a reference to the original movies and these particles would get re-extracted at their original pixel size and then I wouldn’t be able to combine them.

I have thought of some possible work around and I would like to know what the best way to move forward is:

  1. Stay in Cryosparc and process the two particle sets separately, after reference based motion correction I can slightly bin the particles using the downsample particles job to combine the particles.
  2. I can export the project to Relion where I can do Bayesian polishing instead to finish the structure.

What do you think is best? Is there another strategy that I’m not thinking of?

Thank you in advance!

2

Hi,

In this instance, my preference would be to stick with a RELION-centric workflow, mostly because RBMC necessitates Patch Motion Correction input.

If you’re willing to repeat motion correction, e.g. to compare RBMC and Bayesian polishing, then downsampling the eventual output particle stacks to match pixel size sounds reasonable.

Cheers,
Yang

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