Tissue harmonic imaging (THI) has been reported to improve contrast resolution, tissue differentiation and overall image quality in clinical examinations. However, a study carried out previously by the authors (Brown et al. 2004) found improvements only in spatial resolution and not in contrast resolution or anechoic target detection. This result may have been due to the homogeneity of the phantom. Biologic tissues are generally inhomogeneous and THI has been reported to improve image quality in the presence of large amounts of subcutaneous fat. The aims of the study were to simulate the distortion caused by subcutaneous fat to image quality and thus investigate further the improvements reported in anechoic target detection and contrast resolution performance with THI compared with 2D conventional imaging. In addition, the effect of three different types of fat-mimicking layer on image quality was examined. The abdominal transducer of two ultrasound scanners with 2D conventional imaging and THI were tested, the 4C1 (Aspen-Acuson, Siemens Co., CA, USA) and the C5-2 (ATL HDI 5000, ATL/Philips, Amsterdam, The Netherlands). An ex vivo subcutaneous pig fat layer was used to replicate beam distortion and phase aberration seen clinically in the presence of subcutaneous fat. Three different types of fat-mimicking layers (olive oil, lard and lard with fish oil capsules) were evaluated. The subcutaneous pig fat layer demonstrated an improvement in anechoic target detection with THI compared with 2D conventional imaging, but no improvement was demonstrated in contrast resolution performance; a similar result was found in a previous study conducted by this research group (Brown et al. 2004) while using this tissue-mimicking phantom without a fat layer. Similarly, while using the layers of olive oil, lard and lard with fish oil capsules, improvements due to THI were found in anechoic target detection but, again, no improvements were found for contrast resolution for any of the layer combinations. Therefore, it was felt that the lack of improvement in contrast resolution performance may be due to the test phantom design and not to whether a layer was present that caused beam distortion and phase aberrations.