Hi all,

I am a novice at using Relion for Helical Reconstruction. I have a bit of experience with single particle pipeline through the 2D class stage.

I have run one dataset of helical molecules from my own movies images of microtubules through to the 2D class from manual picking, so I know our Relion program is working correctly up to that point. The images I am using were taken on a F20 with K3 camera and they were movie stacks saved as tifs.

I am trying to do the TMV tutorial because I want to understand the finer points (of how asymmetrical units and helical rise parameters in the particle extraction program contribute to the 2D class outcomes), but am stuck at Motion Correction. The tutorial doesn’t not give a specific patch size for the helical images as far as I can find, so I did use the 5 by 5 as recommended for the single particle data set.

ERROR:
Patch size must be even

The images are 3645 by 3750 (x,y) dimensions when I open in Imod

3645/5 = 729, so I guess that is the problem?
3750/5 = 750

I did try 45 by 5 and this did not work either.

I have downloaded the stack.mrc from Empiar 10020, there are 14 images in this tutorial. When I open the files in IMOD, I see there are 22 frames, I am assuming these are the frames from movie stack.

What am I missing? Any help would be greatly appreciated

Thanks for your time,

Cynthia Page

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