Smeared 2D classification?

Hi, all

I had a protein with a molecular weight of 120-kDa. We collected a dataset on K3 camera with the raw images that look good. However, the 2D-classfication results display a smeared pattern after a series of the general processes. I am not sure if the particles locate in the air-water interface or some other mistake happened? Dose someone meet the similar trouble or give any suggestion?



Your 2D look great. There is some flexibility between the two domains (proteins). You should use a slightly smaller box maybe 80% as large, at least until highest resolution reconstructions.

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