The impact of ROI extraction method for MEG connectivity estimation: practical recommendations for the study of resting state data

Magnetoencephalography and electroencephalography (M/EEG) seed-based connectivity analysis requires the extraction of measures from regions of interest (ROI). M/EEG ROI-derived source activity can be treated in different ways. It is possible, for instance, to average each ROI’s time series prior to calculating connectivity measures. Alternatively, one can compute connectivity maps for each element of the ROI prior to dimensionality reduction to obtain a single map. The impact of these different strategies on connectivity results is still unclear. Here, we address this question within a large MEG resting state cohort (N=113) and within simulated data. We consider 68 ROIs (Desikan-Kiliany atlas), two measures of connectivity (phase locking value-PLV, and its imaginary counterpart- ciPLV), three frequency bands (theta 4-8 Hz, alpha 9-12 Hz, beta 15-30 Hz). We compare four extraction methods: (i) mean, or (ii) PCA of the activity within the seed or ROI before computing connectivity, map of the (iii) average, or (iv) maximum connectivity after computing connectivity for each element of the seed. Hierarchical clustering in then applied to compare connectivity outputs across multiple strategies, followed by direct contrasts across extraction methods. Finally, the results are validated by using a set of realistic simulations. We show that ROI-based connectivity maps vary remarkably across strategies in terms of connectivity magnitude and spatial distribution. Dimensionality reduction procedures conducted after computing connectivity are more similar to each-other, while PCA before approach is the most dissimilar to other approaches. Although differences across methods are consistent across frequency bands, they are influenced by the connectivity metric and ROI size. Greater differences were observed for ciPLV than PLV, and in larger ROIs. Realistic simulations confirmed that after aggregation procedures are generally more accurate but have lower specificity (higher rate of false positive connections). Though computationally demanding, after dimensionality reduction strategies should be preferred when higher sensitivity is desired. Given the remarkable differences across aggregation procedures, caution is warranted in comparing results across studies applying different methods. ### Competing Interest Statement The authors have declared no competing interest.