Ultrasonic scattering measurements of a live single cell at 86 MHz

Cell separation and sorting techniques have been employed biomedical applications such as cancer diagnosis and cell gene expression analysis. The capability to accurately measure ultrasonic scattering properties from cells is crucial in making an ultrasonic cell sorter a reality if ultrasound scattering is to be used as the sensing mechanism as well. To assess the performance of sensing and identifying live single cells with high-frequency ultrasound, an 86-MHz lithium niobate pressfocused single-element acoustic transducer was used in a highfrequency ultrasound scattering measurement system that was custom designed and developed for minimizing noise and allowing better mobility. Peak-to-peak echo amplitude, integrated backscatter (IB) coefficient, spectral parameters including spectral slope and intercept, and midband fit from spectral analysis of the backscattered echoes were measured and calculated from a live single cell of two different types on an agar surface: leukemia cells (K562 cells) and red blood cells (RBCs). The amplitudes of echo signals from K562 cells and RBCs were 48.25 ± 11.98 mV<;sub>pp<;/sub> and 56.97 ± 7.53 mV<;sub>pp<;/sub>, respectively. The IB coefficient was -89.39 ± 2.44 dB for K562 cells and -89.00 ± 1.19 dB for RBCs. The spectral slope and intercept were 0.30 ± 0.19 dB/MHz and -56.07 ± 17.17 dB, respectively, for K562 cells and 0.78 ± 0.092 dB/MHz and -98.18 ± 8.80 dB, respectively, for RBCs. Midband fits of K562 cells and RBCs were -31.02 ± 3.04 dB and -33.51 ± 1.55 dB, respectively. Acoustic cellular discrimination via these parameters was tested by Student's t-test. Their values, except for the IB value, showed statistically significant difference (p <; 0.001). This paper reports for the first time that ultrasonic scattering measurements can be made on a live single cell with a highly focused high-frequency ultrasound microbeam at 86 MHz. These results also suggest the feasibility of ultrasonic scattering as a sensing mechanism in the development of ultrasonic cell sorters.