I have a pretty simple question. I ran some initial 2D classifications in Relion and imported my particles to cryosparc for ab initio reconstruction. I notice during import that particle orientations are ignored since I wasn’t importing particles after gold standard refinement and I do not have a random subset assignment for each particle.
I am wondering, since I did a large offset search in relion during 2D classification to find the center of my particles as I was not confident particle picking picked the center of every particle, would it be helpful to do a 2D classification in cryosparc to refine particle offsets before going to ab initio? Or does ab initio not take any particle orientation/offset values anyway?
Thanks in advance!