ObjectivesTo compare the accuracy of pre-surgical prostate size measurements using mpMRI and USWE with imaging-based 3D-printed patient-specific whole-mount moulds facilitated histopathology, and to assess whether size assessment varies between clinically significant and non-significant cancerous lesions including their locations in different zones of the prostate.MethodsThe study population included 202 men with clinically localised prostate cancer opting for radical surgery derived from two prospective studies. Protocol-based imaging data was used for measurement of size of prostate cancer in clinically localised disease using MRI (N = 106; USWE (N = 96). Forty-eight men overlapped between two studies and formed the validation cohort. The primary outcome of this study was to assess the accuracy of pre-surgical prostate cancerous size measurements using mpMRI and USWE with imaging-based 3D-printed patient-specific whole-mount moulds facilitated histopathology as a reference standard. Independent-samples T-tests were used for the continuous variables and a nonparametric Mann–Whitney U test for independent samples was applied to examine the distribution and median differences between mpMRI and USWE groups.ResultsA significant number of men had underestimation of prostate cancer using both mpMRI (82.1%; 87/106) and USWE (64.6%; 62/96). On average, tumour size was underestimated by a median size of 7 mm in mpMRI, and 1 mm in USWE. There were 327 cancerous lesions (153 with mpMRI and 174 for USWE). mpMRI and USWE underestimated the majority of cancerous lesions (108/153; 70.6%) and (88/174; 50.6%), respectively. Validation cohort data confirmed these findings MRI had a nearly 20% higher underestimation rate than USWE (χ2 (1, N = 327) = 13.580, p = 0.001); especially in the mid and apical level of the gland. Clinically non-significant cancers were underestimated in significantly higher numbers in comparison to clinically significant cancers.ConclusionsSize measurement of prostate cancers on preoperative imaging utilising maximum linear extent technique, underestimated the extent of cancer. Further research is needed to confirm our observations using different sequences, methods and approaches for cancer size measurement.