Monitoring of cell death in epithelial cells using high frequency ultrasound spectroscopy

Spectral and wavelet analyses were performed on ultrasound radiofrequency (RF) data collected from centrifuged cell samples containing HEp-2 cells after induction of apoptosis by exposure to camptothecin. Samples were imaged at several time points after drug exposure using high-frequency ultrasound in the range from 10-60 MHz. A 20-MHz transducer with a f-number of 2.35 and a 40-MHz transducer with a f-number of 3 were used for collecting the RF data. Normalized power spectra were computed from the backscattered ultrasound signals within a region-of-interest (ROI) for further analysis. Spectral slopes, integrated backscatter coefficients (IBCs) and wavelet parameters were estimated as a function of treatment time to monitor acoustic property changes during apoptosis. Changes in spectral parameters were detected starting six hours after treatment and coincided with changes in corresponding histology. Throughout the course of chemotherapy, variation in estimates of the spectral slope of up to 35% were observed. During the treatment, IBCs increased by 400% compared with estimates obtained from the control samples. Changes in spectral parameters are hypothesized to be linked to structural cell changes during apoptosis. In addition, the sensitivity of a wavelet-based analysis to the ultrasonic assessment of cellular changes was investigated. Results of the wavelet analysis showed variations similar to the spectral parameters. Where values of the spectral slope decreased, estimates of the scaling factors increased. Because wavelet analysis preserves the signal-time localization, its application will be potentially beneficial for assessing treatment responses in vivo. The current study contributes toward the development of a non-invasive method for monitoring apoptosis as a measure of the success of chemotherapeutic treatment of cancer.