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Hi, In the RELION multi-body paper, it is stated that: “...the relion_flex_analyse program can output subtracted images according to the iteratively refined relative orientations from a multi-body refinement. The resulting subtracted particle images can then be used in subsequent focused classifications and/or refinements. An additional advantage of this program over previously existing subtraction tools in relion is the centering at the projected centre-of-mass of the subtracted particle images, which allows the use of more meaningful priors and smaller box sizes.” Is this option (recentering and reboxing around the center of mass of a subregion) available anywhere else in Relion 3, as an independent job outside of the multi-body module? It would be a very handy option for existing focused refinements, or during signal subtraction. Cheers Oli ######################################################################## To unsubscribe from the CCPEM list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCPEM&A=1