High-intensity focused ultrasound (HIFU) is becoming an increasingly important noninvasive surgical tool, despite the challenges in temperature rise control and unwanted heating problems. In this study, experiments and simulations on tissue ablation effectiveness were performed using multi-frequency HIFU with frequency differences of more than 500 kHz (center frequencies are 950 kHz, 1.5 MHz, and 3.3 MHz). In the experiments, the temperature was recorded as chicken breast tissue was heated by single-frequency, dual-frequency, and tri-frequency HIFU configurations at controlled acoustic power and exposure time. 5% to 10% temperature rise differences were observed between single- and multi-frequency modes, indicating that multi-frequency HIFU is more effective at producing faster temperature rises. Cavitation detection tests were conducted to compare the cavitation pressure fields between single- and multi-frequency ultrasound. Moreover, simulations on single-frequency and multi-frequency acoustic fields as well as bio-heating-induced temperature fields were performed. With the comparison between experimental and simulation results, we believe that the more effective tissue ablation using multi-frequency ultrasound is likely attributed to the enhanced cavitation, a promising result for HIFU applications.