Acoustofluidic devices becomes one of the emerging and versatile tools for many biomedical applications. Most of the previous acoustofluidic devices are used for cells manipulation, and the few devices for cell phenotyping with a limitation in throughput. In this study, an enhanced tilted-angle (ETA) acoustofluidic device is developed and applied for mechanophenotyping of live cells. The ETA Device consists of an interdigital transducer which is positioned along a microfluidic channel. An inclination angle of 5° is introduced between the interdigital transducer and the liquid flow direction. The pressure nodes formed inside the acoustofluidic field in the channel deflect the biological cells from their original course in accordance with their mechanical properties, including volume, compressibility, and density. The threshold power for fully converging the cells to the pressure node is used to calculate the acoustic contrast factor. To demonstrate the ETA device in cell mechanophenotyping, and distinguishing between different cell types, further experimentation is carried out by using A549 (lung cancer cells), MDB-MA-231 (breast cancer cells), and leukocytes. The resulting acoustic contrast factors for the lung and breast cancer cells are different from that of the leukocytes by 27.9% and 21.5%, respectively. These results suggest this methodology can successfully distinguish and phenotype different cell types based on the acoustic contrast factor.