Laser interstitial thermotherapy (LITT) is a relatively new focal therapy technique for the ablation of localized prostate cancer. However, very little is known about the specific effects of LITT within the ablation zone and the surrounding normal tissue regions. For instance, it is important to be able to assess the extent of residual cancer within the prostate following LITT, which may be masked by thermally induced benign necrotic changes. Fortunately LITT is MRI compatible and hence this allows for quantitatively assessing LITT induced changes via multi-parametric MRI. Of course definite validation of any LITT induced changes on MRI requires confirmation via histopathology. The aim of this study was to quantitatively assess and distinguish the imaging characteristics of prostate cancer and benign confounding treatment changes following LITTon 3 Tesla multi-parametric MRI by carefully mapping the treatment related changes from the ex vivo surgically resected histopathologic specimens onto the pre-operative in vivo imaging. A better understanding of the imaging characteristics of residual disease and successfully ablated tissue might lead to improved treatment monitoring and as such patient prognosis. A unique clinical trial at the Radboud University Medical Center, in which 3 patients underwent a prostatectomy after LITT treatment, yielded ex-vivo histopathologic specimens along with pre- and post-LITT MRI. Using this data we (1) identified the computer extracted MRI signatures associated with treatment effects including benign necrotic changes and residual disease and (2) subsequently evaluated the computer extracted MRI features previously identified in distinguishing LITT induced changes in the ablated area relative to the residual disease. Towards this end first a pathologist annotated the ablated area and the residual disease on the ex-vivo histology and then we transferred the annotations to the post-LITT MRI using semi-automatic elastic registration. The pre- and post-LITT MRI were subsequently registered and computer-derived multi-parametric MRI features extracted to determine differences in feature values between residual disease and successfully ablated tissue to assess treatment response. A scoring metric allowed us to identify those specific computer-extracted MRI features that maximally and differentially expressed between the ablated regions and the residual cancer, on a voxel- by voxel basis. Finally, we used a Fuzzy C-Means algorithm to assess the discriminatory power of these selected features. Our results show that specific computer-extracted features from multi-parametric MRI differentially express within the ablated and residual cancer regions, as evidenced by our ability to, on a voxel-by-voxel basis, classify tissue as residual disease. Additionally, we show that change of feature values between pre- and post-LITT MRI may be useful as a quantitative marker for treatment response (T2-weighted texture and DCE MRI features showed largest differences between residual disease and successfully ablated tissue). Finally, a clustering approach to separate treatment effects and residual disease incorporating both (1) and (2) yielded a maximum area under the ROC curve of 0.97 on a voxel basis across 3 studies.