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Hi all, I'm running the 3D auto-refine for my data and it shows the WARNING about the angular accuracy is worse than 10 degrees. I'm just wondering: 1. What cause the accuracy worse than 10˚? 2. I checked the map for iteration6, even the shape is not correct. Should I stop the job? 3. To tune resolution by adjusting T, should I run 3D-classification after refinement or skip refinement just run 3D-classification with a single class. Thanks, Fuxing Auto-refine: Resolution= 38.176 (no gain for 2 iter) Auto-refine: Changes in angles= 29.0102 degrees; and in offsets= 1.14429 pixels (no gain for 0 iter) Estimating accuracies in the orientational assignment ... 4/ 4 sec ............................................................~~(,_,"> Auto-refine: Estimated accuracy angles= 30.1 degrees; offsets= 10.1 pixels Auto-refine: WARNING: The angular accuracy is worse than 10 degrees, so basically you cannot align your particles (yet)! Auto-refine: WARNING: You probably need not worry if the accuracy improves during the next few iterations. Auto-refine: WARNING: However, if the problem persists it may lead to spurious FSC curves, so be wary of inflated resolution estimates... Auto-refine: WARNING: Sometimes it is better to tune resolution yourself by adjusting T in a 3D-classification with a single class. Auto-refine: Angular step= 7.5 degrees; local searches= false Auto-refine: Offset search range= 5 pixels; offset step= 1 pixels CurrentResolution= 38.176 Angstroms, which requires orientationSampling of at least 12.8571 degrees for a particle of diameter 340 Angstroms Oversampling= 0 NrHiddenVariableSamplingPoints= 96768 OrientationalSampling= 15 NrOrientations= 4608 TranslationalSampling= 2 NrTranslations= 21 ============================= Oversampling= 1 NrHiddenVariableSamplingPoints= 3096576 OrientationalSampling= 7.5 NrOrientations= 36864 TranslationalSampling= 1 NrTranslations= 84 ============================= Estimated memory for expectation step > 0.318263 Gb, available memory = 40 Gb. Estimated memory for maximization step > 0.0255431 Gb, available memory = 40 Gb.