A cell pellet biophantom technique is introduced, and applied to the ultrasonic backscatter coefficient (BSC) estimate using Chinese hamster ovary (CHO) cells. Also introduced is a concentric sphere scattering model because of its geometrical similarities to cells with a nucleus. BSC comparisons were made between the concentric sphere model and other well-understood models for mathematical verification purposes. BSC estimates from CHO cell pellet biophantoms of known number density were performed with 40 and 80 MHz focused transducers (overall bandwidth: 26-105 MHz). These biophantoms were histologically processed and then evaluated for cell viability. Cell pellet BSC estimates were in agreement with the concentric sphere model. Fitting the model to the BSC data yielded quantitative values for the outer sphere and inner sphere. The radius of the cell model was 6.8 ± 0.7 μm; the impedance of the cytoplasm model was 1.63 ± 0.03 Mrayl and the impedance of the nuclear model was 1.55 ± 0.09 Mrayl. The concentric sphere model appears as a new tool for providing quantitative information on cell structures and will tend to have a fundamental role in the classification of biological tissues.

Simple scattering media fit scattering model theories much better than more complex scattering media. Tissue is much more complex as an acoustic scattering media and to date there has not been an adequate scattering model that fits it well. Previous studies evaluated the scattering characteristics of simple media (grouping of cells at various number densities) and fit them to the concentric spheres scattering model theory. This study is to increase the complexity of the media to provide insight into the acoustic scattering characteristics of tissue, and specifically two tumor types. Complementing the data from the tumors is 100% volume fraction cell pellets of the same cell lines. Cell pellets and ex vivo tumors are scanned using high-frequency single-element transducers (9-105 MHz), and the attenuation and backscatter coefficient (BSC) are estimated. BSC comparisons are made between cell pellets and tumors. The results show that the 4T1 (ATCC #CRL-2539) cell pellets and tumors have similar BSC characteristics, whereas the MAT (ATCC #CRL-1666) cell pellets and tumors have significantly different BSC characteristics. Factors that yield such differences are explored. Also, the fluid-filled sphere and the concentric spheres models are evaluated against the BSC characteristics, demonstrating that further work is required.

Previous work estimated the ultrasonic backscatter coefficient (BSC) from low-concentration (volume density <3%) Chinese Hamster Ovary (CHO, 6.7-μm cell radius) cell pellets. This study extends the work to higher cell concentrations (volume densities: 9.6% to 63%). At low concentration, BSC magnitude is proportional to the cell concentration and BSC frequency dependency is independent of cell concentration. At high cell concentration, BSC magnitude is not proportional to cell concentration and BSC frequency dependency is dependent on cell concentration. This transition occurs when the volume density reaches between 10% and 30%. Under high cell concentration conditions, the BSC magnitude increases slower than proportionally with the number density at low frequencies (ka<1), as observed by others. However, what is new is that the BSC magnitude can increase either slower or faster than proportionally with number density at high frequencies (ka>1). The concentric sphere model least squares estimates show a decrease in estimated cell radius with number density, suggesting that the concentric spheres model is becoming less applicable as concentration increases because the estimated cell radius becomes smaller than that measured. The critical volume density, starting from when the model becomes less applicable, is estimated to be between 10% and 30% cell volume density.

Quantitative ultrasound (QUS) imaging is hypothesized to map temperature elevations induced in tissue with high spatial and temporal resolution. To test this hypothesis, QUS techniques were examined to monitor high-intensity focused ultrasound (HIFU) exposure of tissue. In situ experiments were conducted on mammary adenocarcinoma tumors grown in rats and lesions were formed using a HIFU system. A thermocouple was inserted into the tumor to provide estimates of temperature at one location. Backscattered time-domain waveforms from the tissue during exposure were recorded using a clinical ultrasonic imaging system. Backscatter coefficients were estimated using a reference phantom technique. Two parameters were estimated from the backscatter coefficient (effective scatterer diameter (ESD) and effective acoustic concentration (EAC). The changes in the average parameters in the regions corresponding to the HIFU focus over time were correlated to the temperature readings from the thermocouple. The changes in the EAC parameter were consistently correlated to temperature during both heating and cooling of the tumors. The changes in the ESD did not have a consistent trend with temperature. The mean ESD and EAC before exposure were 120 ± 16 μm and 32 ± 3 db/cm3, respectively, and changed to 144 ± 9 μm and 51 ± 7 db/cm3, respectively, just before the last HIFU pulse was delivered to the tissue. After the tissue cooled down to 37°C, the mean ESD and EAC were 126 ± 8 μm and 35 ± 4 db/cm3, respectively. Peak temperature in the range of 50–60°C was recorded by a thermocouple placed just behind the tumor. These results suggest that QUS techniques have the potential to be used for non-invasive monitoring of HIFU exposure.