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Objectives To compare elastography measurements in men with normal testicular tissue, testicular microlithiasis and testicular cancer. Methods A total of 248 consecutive patients were included. All men provided written informed consent. Testicular stiffness was assessed using shear wave elastography (SWE). Three SWE velocity measurements were assessed in each testicle. The patients were divided into three groups; men with normal testicular tissue (n=130), men with testicular microlithiasis (n=99) and men with testicular cancer (n=19). Results We found a higher mean velocity in the group of patients with testicular cancer (1.92 m/s (95% CI 1.82–2.03)) compared to both the group with normal tissue (0.76 m/s (95% CI: 0.75–0.78)) (p<0.001) and the group with testicular microlithiasis 0.79 m/s (95% CI: 0.77–0.81) (p<0.001). Conclusion The presence of testicular microlithiasis increased stiffness slightly, but within the range of variation in normal testicles. Increased stiffness may indicate testicular malignancy in testicular lesions. Ultrasound elastography could be a very useful tool when investigating scrotum.