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Dear Chia, The pixel size on the GUI will be internally corrected. Changing it on the GUI is not strictly necesssary, but will prevent warnings in the refinement runs. The important thing is that the physical detector pixel size in the input data.star file is now 5x larger. Also, if you want to re-use translations (rlnOriginX/Y) from a previous refinement, please be aware that those are in pixels, so you would need to divide those columns by 5. awk is a great tool for that. HTH, Sjors > Hi everyone > > I processed my unbinned data until particle extraction step. > Then I decide to do a binning of 5, due to the limitation of computation > resource. > > The magnified pixel size in the GUI was 1.06 and the particle mask > diameter was 320. > so, I set Re-scaled size (pixels) in the GUI to 64. > > In order to do particle sorting, I also rescale my reference by > relion_image_handler with > "--angpix 1.06 --rescale_angpix 5.30 --new_box 64" > > My questions are: > 1.) am I on the right track so far ? > 2.) after all these steps, in the gui should I changed the The magnified > pixel size in the GUI to 5.3 or leave it as 1.06? > I could do particle sorting in both cases. The two sortings look a > bit different but both seems ok. > > Thanks. > > Chia > -- Sjors Scheres MRC Laboratory of Molecular Biology Francis Crick Avenue, Cambridge Biomedical Campus Cambridge CB2 0QH, U.K. tel: +44 (0)1223 267061 http://www2.mrc-lmb.cam.ac.uk/groups/scheres