Motion corrected micrographs from Relion 3.1.3

[forhad]

Hi there,
We are analysing some data collected through EPU group AFIS strategy using a Falcon 4 detector (EERs). I have created the optics.star file and did Motion Correction on Relion 3.1.3. Then I imported the Motion Corrected .mrc micrographs to CS. I labelled the Relion’s Motion Corrected Micrographs’ directory as the movie path and used the *.mrc wild card. Then I did Patch CTF Estimation (multi) on CS and keep processing as usual. Is that a reasonable approach with this EPU_AFIS strategy?

During Inspection of Blob picks, I noticed the power threshold went only upto 140. Also, here are 2 examples of CTF estimation with CS and Relion (CTFFind), respectively:

We are still using CS 3.3.1. Experts, please suggest.

Regards,
Forhad, UQ.

[olibclarke]

Hi Forhad,

Yes, this is a reasonable strategy.

The display of the power spectrum on motion corrected mics is some kind of display bug, I reported it a while back but it doesn’t seem to affect the quality of the underlying CTF fits (just display of the PS). What do the 1D plots & estimated CTF fit resolutions look like?

Cheers
Oli

[forhad]

Hi @olibclarke,

Thanks for your reply. This is one of the representative 1D plots:

This is the overview of CTF fit res. During Manually Curate Exposure job, I limited them by roughly 10 angs.

Any critical comment?

Regards,
Forhad.

[olibclarke]

they look on the lower res side, but not inconsistent with the PS you showed from CTFFIND. I would select and double check the ones with estimated fits of >10Å, and see why they are failing.

[forhad]

Hi @olibclarke,

Here is an example of a mic with an estimated fit of 33.72 angs.