I’m using cryosparc live to import .eer files collected from falcon 4 camera and pick particles.
Although the images are in .eer format, the gain reference file is in .mrc format.
I’m using upsampling factor of 2 and 40 frames.
I have processed the same data set twice, once with the gain reference and once without. The reconstruction calculated from images without the gain reference applied achieved a higher resolution by 0.2 Å, indicating that the reference had been applied incorrectly. The gain reference file has already been accounted for flip or rotation, so I don’t think that would have been the problem.
What is the correct way to apply .mrc gain reference file for .eer file?
It seemed like other tools (ex. Relion) requires multiplying or dividing pixel values of the gain reference file depending on the upsampling factor. Does Crysparc also require something like that too?
Thanks in advance for your help!