Skip to content
New issue

Have a question about this project? Sign up for a free GitHub account to open an issue and contact its maintainers and the community.

By clicking “Sign up for GitHub”, you agree to our terms of service and privacy statement. We’ll occasionally send you account related emails.

Already on GitHub? Sign in to your account

Preprocessing::initialise ERROR: Input micrograph STAR file has no rlnMicrographName column! #773

Closed
yongshuo-Z opened this issue May 12, 2021 · 12 comments
Labels

Comments

@yongshuo-Z
Copy link

Describe your problem
After importing raw micrographs and its corresponding coordinates, I tried to extract particles. But error occurs as the issue title. But I tried to read the .star file using python myself, I found the exact rlnMicrographName as below. I wonder what should I do to extract particles?

image

Environment:

  • OS: Ubuntu 18.04 LTS
  • RELION version RELION-3.1.2-commit-dcab79 # also tried version 2.1.1beta, same error occurs
  • Memory: 128 GB
  • GPU: GTX 2080Ti

Dataset:

  • Box size: [e.g. 256 px]
  • Pixel size: [e.g. 0.9 Å/px]
  • Number of particles: [e.g. 150,000]
  • Description: [e.g. A tetrameric protein of about 400 kDa in total]

Job options:

  • Type of job: Particle Extraction

  • Number of MPI processes: 1

  • Full command (see note.txt in the job directory):

    
    

relion_import --do_movies --optics_group_name "opticsGroup1" --angpix 1.4 --kV 300 --Cs 2.7 --Q0 0.1 --beamtilt_x 0 --beamtilt_y 0 --i "data/empiar10017/*.mrc" --odir Import/job001/ --ofile movies.star --continue --pipeline_control Import/job001/

relion_import --do_coordinates --i "data/empiar10017/*_checked.star" --odir Import/job002/ --ofile coords_suffix_checked.star --continue --pipeline_control Import/job002/

relion_preprocess` --i Import/job001/movies.star --coord_dir Import/job002/ --coord_suffix _checked.star --part_star Extract/job003/particles.star --part_dir Extract/job003/ --extract --extract_size 128 --norm --bg_radius 48 --white_dust -1 --black_dust -1 --invert_contrast --only_do_unfinished --pipeline_control Extract/job003/


**Error message:**

in: /root/cryo/code/relion/relion/src/preprocessing.cpp, line 138
ERROR:
Preprocessing::initialise ERROR: Input micrograph STAR file has no rlnMicrographName column!
=== Backtrace ===
./relion_preprocess(_ZN11RelionErrorC2ERKNSt7__cxx1112basic_stringIcSt11char_traitsIcESaIcEEES7_l+0x77) [0x7f8dc2d4d7a7]
./relion_preprocess(_ZN13Preprocessing10initialiseEv+0xcfe) [0x7f8dc2d70b8e]
./relion_preprocess(main+0x49) [0x7f8dc2d422f9]
/lib/x86_64-linux-gnu/libc.so.6(__libc_start_main+0xe7) [0x7f8db83afbf7]
./relion_preprocess(_start+0x2a) [0x7f8dc2d453da]

ERROR:
Preprocessing::initialise ERROR: Input micrograph STAR file has no rlnMicrographName column!

@biochem-fan
Copy link
Member

You cannot extract from movies. Run motion correction first.

@njxisang
Copy link

How do you solve it? I have met the same error after I use the motion correction.

@biochem-fan
Copy link
Member

Did you run motion correction inside RELION?

@njxisang
Copy link

Did you run motion correction inside RELION?

Yes, I have done it, I get the corrected_micrographs.star and then I use the ctf estimation based on corrected_micrographs.star to get the micrographs_ctf.star.
The micrographs_ctf.star is as follows and I use Relion-3.1.3

version 30001

data_optics

loop_
_rlnOpticsGroupName #1
_rlnOpticsGroup #2
_rlnMicrographPixelSize #3
_rlnMicrographOriginalPixelSize #4
_rlnVoltage #5
_rlnSphericalAberration #6
_rlnAmplitudeContrast #7
opticsGroup1 1 1.400000 1.400000 300.000000 2.700000 0.100000

@biochem-fan
Copy link
Member

The micrographs_ctf.star is as follows and I use Relion-3.1.3

Is this all? Don't you have the data_micrographs table after what yo have cited?

@njxisang
Copy link

I am not very familiar with this. I use this MRC file (stack_0220_2x_SumCorr.mrc) as the single particle extraction. By import, correction and ctf, I get the file micrographs_ctf.star:
version 30001
data_optics

loop_
_rlnOpticsGroupName #1
_rlnOpticsGroup #2
_rlnMicrographPixelSize #3
_rlnMicrographOriginalPixelSize #4
_rlnVoltage #5
_rlnSphericalAberration #6
_rlnAmplitudeContrast #7
opticsGroup1 1 1.400000 1.400000 300.000000 2.700000 0.100000

Yes, it is all I get.

@biochem-fan
Copy link
Member

biochem-fan commented Jun 16, 2022

Is stack_0220_2x_SumCorr.mrc the original file you started with?
The file name suggests that your inputs are not movies but motion corrected micrographs.

So "Did you run motion correction inside RELION?" "Yes, I have done it" is not correct.
You cannot perform motion correction on non-movies!

Import these files as micrographs, not movies, and then perform CTF estimation and extraction.

@njxisang
Copy link

Is stack_0220_2x_SumCorr.mrc the original file you started with? The file name suggests that your inputs are not movies but motion corrected micrographs.

So "Did you run motion correction inside RELION?" "Yes, I have done it" is not correct. You cannot perform motion correction on non-movies!

Import these files as micrographs, not movies, and then perform CTF estimation and extraction.

Yes, I have imported it as micrographs and the perform CTF estimation. Then, I get micrographs_ctf.star as follows:

version 30001

data_optics

loop_
_rlnOpticsGroupName #1
_rlnOpticsGroup #2
_rlnMtfFileName #3
_rlnMicrographPixelSize #4
_rlnMicrographOriginalPixelSize #5
_rlnVoltage #6
_rlnSphericalAberration #7
_rlnAmplitudeContrast #8
opticsGroup1 1 stack_0220_2x_SumCorr.mrc 1.400000 1.400000 300.000000 2.700000 0.100000

However, when I use it to extraction, it also report the bug:
Preprocessing::initialise ERROR: Input micrograph STAR file has no rlnMicrographName column!

The imported file I get is micrographs.star as follows:

version 30001

data_optics

loop_
_rlnOpticsGroupName #1
_rlnOpticsGroup #2
_rlnMtfFileName #3
_rlnMicrographPixelSize #4
_rlnMicrographOriginalPixelSize #5
_rlnVoltage #6
_rlnSphericalAberration #7
_rlnAmplitudeContrast #8
opticsGroup1 1 stack_0220_2x_SumCorr.mrc 1.400000 1.400000 300.000000 2.700000 0.100000

@biochem-fan
Copy link
Member

You are importing in a wrong way. Why did you specify the image as an MTF file?

If this is your first time using RELION, please practice on our tutorial first.

@njxisang
Copy link

You are importing in a wrong way. Why did you specify the image as an MTF file?

If this is your first time using RELION, please practice on our tutorial first.

Oh, I am sorry. However, it is my first time to use Relion. However, the first I import the file is not MTF file and I get the file is:

version 30001

data_optics

loop_
_rlnOpticsGroupName #1
_rlnOpticsGroup #2
_rlnMicrographPixelSize #3
_rlnMicrographOriginalPixelSize #4
_rlnVoltage #5
_rlnSphericalAberration #6
_rlnAmplitudeContrast #7
opticsGroup1 1 1.400000 1.400000 300.000000 2.700000 0.100000

version 30001

data_micrographs

loop_
_rlnMicrographName #1
_rlnOpticsGroup #2
stack_0220_2x_SumCorr.mrc 1

But it also reports the same bug. Thank you very much for your kind reply.

@biochem-fan
Copy link
Member

It is getting better. Now you have the data_micrographs table.
But why do you have only one file? Did you use the wild card?

However, it is my first time to use Relion.

PLEASE go through the tutorial first! Since this is not a RELION bug but your wrong usage, I stop answering this issue here.

If the problem persists even after you learn the tutorial, please ask in the CCPEM mailing list.

@njxisang
Copy link

I am so sorry to bother you. I will learn it carefully, thank you for kind reply again.

Sign up for free to join this conversation on GitHub. Already have an account? Sign in to comment
Labels
Projects
None yet
Development

No branches or pull requests

3 participants