The authors have proposed a new type of ultrasonic microscopy for biological tissue characterization. The system is driven by a nanosecond pulse voltage, the generated acoustic wave being reflected at the front and rear side of the sliced tissue. In this report, a time-frequency analysis was applied to determine the sound speed thorough the tissue. Frequency dependence of sound speed was obtained with a myocardium of a rat sliced into 10 μm. As the reflected waveform had a significant amount of oscillating component, the waveform was once subjected to the deconvolution process. As the result, two reflections were clearly separated in time domain. Then these two reflections were separately analyzed by time-frequency analysis. Each reflection was extracted by using a proper window function. Phase angles of these reflections at the same frequency were compared. A sound speed micrograph at an arbitrary frequency in between 50 and 150 MHz was successfully obtained. A tendency was found that the sound speed slightly increases with frequency.
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