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Hello, (1) The Colin27 is the brain of a single subject, and cannot provide probability maps as needed by SPM5 for segmentation. So unless there is a very specific reason to no use the default priors in the tmp folder, these default option would be the first solution. (2) No. Unified segmentation runs via the segmentation button and works with three tissue priors. The normalize button - to my knowledge - is about doing the same as compared with normalization in SPM2. In both versions you can either estimate the deformation matrix or estimate and write out the normalized brains. Hope this helps - if not, best explain what the goal of the SPM2/SPM5 comparison is, there have been mails on the differences between these two versions, Philipp Max Planck Institute of Psychiatry NMR Research Group Kraepelinstr. 2-10 80804 Munich Mail: [log in to unmask] Phone: 0049-89-30622-413