I am a beginner in Cryo EM. I have collected images of my protein which is about 150kDa and has a diameter of about 180A. While doing 2D classificiation, I am getting very poor classes. I have extracted the particles using box size 384→96. I am also attaching the 2D classification parameters for your reference.
Number of 2D classes: 200
Maximum resolution (A): 8
Force max over poses/shifts: Off
Number of final full iterations: 2
Number of online-EM iterations: 40
Batchsize per class: 200
Enforce non-negativity: On
Use clamp-solvent to solve 2D classes: On
Sort classes by number of particles: Off
Hard classify for last iteration: On
Thanks is advance
Samrat