Far field during sonication experiments in vitro - Is it really far enough?

In many in vitro experiments studying ultrasound bioeffects the sonicated samples are placed to far field with intention of exposing them to as uniform ultrasound field as possible. The main aim of this work is to assess whether the sonicated samples really experience what they are believed to. Also we would like to suggest basic rules for construction of sonication vessels. We used 3.5 MHz and 7 MHz ultrasound transducers for measurements. We measured ultrasound field inside and behind common culture plates and special 3D printed plates placed to last axial maximum in water sonication tank with use of a needle hydrophone. Our results show that even though the sonication vessels with sonicated samples are placed into far field, the sonicated samples are actually exposed to some kind of a near field pattern which develops due to the interaction between ultrasound and well of culture plate. The variability of local acoustic intensity can reach up to several hundreds of percent. Our results are also supported by theoretical calculation and software for simulation of ultrasound fields. Even though the sonicated samples may have actually been exposed to some kind of near field pattern in many past studies, the whole phenomenon of creation of near field pattern can be controlled to some extent for future studies. Thus, we suggest that the sonication vessel should always be designed for particular ultrasound transducer.