Background subtraction after Patch Motion Correction

Motion Correction
1

Hello,

I am trying Strep-Affinity grids for my project (following this protocol Streptavidin-Affinity Grid Fabrication for Cryo-Electron Microscopy Sample Preparation (Scientific Article Protocol) | JoVE ).

I need to subtract the strep background from the motion corrected micrographs. I was thinking of using Patch Motion Correction job → take the corrected micrographs → subtract the background → feed the subtracted micrographs back to cryoSPARC and do CTF est. etc.

I am bit confused which file to use though. As far as I understand the outputs, CS uses …_patch_aligned.mrc files for the CTF estimation, but down the line uses DW …_patch_aligned_doseweighted.mrc micrographs to extract the particles?

So which file would be the one for the background subtraction? Should I just use the DW file?

Thanks,

Marek

2

Hi @MmM,

For this workflow, you will want to do the following:

  1. Patch motion
  2. Subtract out streptavidin using the doseweighted mics
  3. Import the streptavidin-subtracted mics into CS
  4. Perform Patch CTF estimation

Mics will be usable for any downstream processing, except for reference based motion correction.

Cheers,
Kye