Blood brain barrier (BBB) opening by MR-guided pulsed Focused Ultrasound (pFUS) and microbubbles (MB) is a non-invasive treatment of various central nervous system diseases. However, the potential adverse effects of repeated pFUS+MB exposure have not been thoroughly elucidated and may limit clinical translation. To date MRI scans of repeated BBB opening by pFUS+MB have been achieved without hemorrhage, edema and behavioral changes in non-human primates (Arvanitis, et al. 2015; Downs, et al. 2015). By incorporating detailed multivariate imaging modalities we characterized the long term effects of single or repeated pFUS+MB in the rat brain.