Measurement of ultrasound velocity in yolk and blastula of fish embryo in vivo

Abstract

An acoustic microscopy method for measuring the velocity of ultrasound in the yolk and blastula of bony fish embryos at early stages of development was proposed. The yolk and blastula were approximated as a sphere and a spherical dome, respectively, consisting of a homogeneous liquid. A theoretical model of ultrasonic wave propagation through a spherical liquid drop located on a solid substrate was developed in the ray approximation. The dependence of the wave propagation time on the speed of sound in the drop, its diameter, and the position of the focus of the ultrasonic transducer has been determined. It was shown that the velocity in the drop can be found by solving the inverse problem by minimizing the discrepancy between the experimental and model spatial distributions of the propagation time, assuming that the velocity in the immersion liquid and the radius of the drop are known. The velocities in the yolk and blastula of the loach (Misgurnus fossilis) embryo at the stage of development of the middle blastula were measured in vivo using a pulsed scanning acoustic microscope operating at a central frequency of 50 MHz. The yolk and blastula radii were determined from ultrasound images of the embryo. Acoustic microscopy measurements conducted with four embryos provide velocities of the acoustic longitudinal wave in the yolk and blastula. They were measured to be 1581 ± 5 m/s and 1525 ± 4 m/s when the temperature of the liquid in the water tank was kept at 22 ± 2 °C.