I've had the same issue and haven't found a setting that could correct for this.
It seems that Relion is a lot more sensitive to the ribosome conformation than it is to tRNA occupancies.
The only solution I see is to place a mask around the tRNA's and classify with very restricted angles. Of course the particles should be aligned first.
Amedee des Georges
Howard Hughes Medical Institute
Dept. of Biochemistry and Molecular Biophysics
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New York, NY 10032
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On Aug 9, 2013, at 3:08 PM, Zhu Hongtao <[log in to unmask]> wrote:
> I have ran my ribosome data in relion-1.2. After 3d classification, I could always get the state with three tRNA inside. And that must contain two sub states at least. But no matter how many class numbers I set, I can not separate them out. Could you give me some suggestions? Thanks!