Relion polish Error

Troubleshooting
1

Hi,
I am trying to use relion to do polish, and after running it, an error message appears.
error: not a single movie contains the requested number of frames (32).
In the run.err, it also shows that Movies for the following micrographs are missing: Motioncorr/iob003/RawTif/stack 00001.mrc.
However, in the Motioncorr/iob003/RawTif/ folder, the stack 00001.mrc file is confirmed to exist.
I would like to ask how to solve this error?

Thanks!

2

mrc, shouldn’t able to do motion correct

3

MRC is perfectly able to be motion corrected if it is a multi-frame stack. EPU used to save micrograph movies in MRC format (and still can). As can SerialEM etc.

It’s more likely caused by the apparent space in the filename, and RELION not coping well with directories or filenames with spaces in.

1 Like
4

thanks
The space was accidentally ignored when I posted a question on the website. In fact, the file name contains underscores and should be stack_00001.mrc.
I have also posted the picture of the error report.

1
1908×950 189 KB

5

2
2844×600 212 KB

6

But Error should indicate that this MRC file is not a movie?

7

To me this does not look like the micrographs in ‘MotionCorr/job003/RawTif/…’ are missing, but the raw data that is associated with the motion corrected micrographs. In the MotionCorr folder you find the motioncorrected micrographs, not the movies. Judging from the folder structure, relion expects them to be in a folder called ‘RawTif’ in your processing folder. Can you double check whether your raw data movies are actually there?

8

Thanks for your advice
I’ve checked this multiple times. In the MotionCorr/job003/RawTif/ folder, stack_???.mrc does exist.
So I’m really confused

9

You are talking about the Motion corrected micrographs. Relion is looking for the raw data movies, which are located in a folder called /RawTiff in you processing folder. Can you make sure that you have a folder with Tiff files?

10

Thanks
The problem has been solved
These errors occur because there is no RawTif folder in my processing folder. Because I only copied the MotionCorr folder, but did not copy the folder where the original movie is located (RawTif), polish needs to call the original movie.
When I copy the RawTif folder containing the movie, it can run normally.