Group analysis and the subject factor in functional magnetic resonance imaging: Analysis of fifty right-handed healthy subjects in a semantic language task

Before considering a given fMRI paradigm as a valid clinical tool, one should first assess the reliability of functional responses across subjects by establishing a normative database and defining a reference activation map that identifies major brain regions involved in the task at hand. However, the definition of such a reference map can be hindered by inter-individual functional variability. In this study, we analysed functional data obtained from 50 healthy subjects during a semantic language task to assess the influence of the number of subjects on the reference map and to characterise inter-individual functional variability. We first compared different group analysis approaches and showed that the extent of the activated network depends not only on the choice of the analysis approach but also on the statistical threshold used and the number of subjects included. This analysis suggested that, while the RFX analysis is suitable to detect confidently true positive activations, the other group approaches are useful for exploratory investigations in small samples. The application of quantitative measures at the voxel and regional levels suggested that while similar to 15-20 subjects were sufficient to reveal reliable and robust left hemisphere activations, >30 subjects were necessary for revealing more variable and weak right hemisphere ones. Finally, to visualise inter-individual variability, we combined two similarity indices that assess the percentages of true positive and false negative voxels in individual activation patterns relative to the group map. We suggest that these measures can be used for the estimation of the degree of 'normality' of functional responses in brain-damaged patients, where this question is often raised, and recommend the use of different quantifications to appreciate accurately the inter-individual functional variability that can be incorporated in group maps.