Quantitative analysis of the angiogenic microvasculature in tumor using multiple scattering and dual-frequency transducers

The objective is to characterize angiogenic networks using contrast-enhanced multiple scattering. The methods relies on the measurement of the diffusion constant from the time evolution of the incoherent backscattered intensity. Dual-frequency arrays allow to emit a pulse at a frequency close to resonance frequency of microbubbles, and to receive the backscattered echoes at higher harmonics, resulting in better contrast-to-tissue ratio compared to single frequency transducers. At 8 MHz, we demonstrated that the diffusion constant enables the quantification of the vascular density and anisotropy in tumor-related vasculature in rat models in vivo. We show here that the results could be further improved using a dual-frequency approach, using two linear arrays with different central frequencies (7 MHz/18 MHz) and using a custom made dual frequency transducer (3 MHz/15 MHz). The minimum detectable distance between two vessel-mimicking cellulose tubes filled with contrast agents was measured via the diffusion constant. The dual-frequency approach drastically reduced the smallest detectable distance between tubes, which was found to be 25µm using a single frequency, 20µm using two linear arrays and 10µm using the custom dual-frequency array, respectively. These results show that measuring the diffusion constant with a dual frequency array can enable the quantification of vessel density with high accuracy