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Acoustic levitation provides potential to characterize and manipulate material such as solid particles and fluid in a wall-less environment. While attempts to levitate small animals have been made, the biological effects of such levitation have been scarcely documented. Here, our goal was to explore if zebrafish embryos can be levitated (peak pressures at the pressure node and anti-node: 135 dB and 144 dB, respectively) with no effects on early development. We levitated the embryos ( n  = 94) at 2–14 hours post fertilization (hpf) for 1000 ( n  = 47) or 2000 seconds ( n  = 47). We compared the size and number of trunk neuromasts and otoliths in sonicated samples to controls ( n  = 94), and found no statistically significant differences ( p  > 0.05). While mortality rate was lower in the control group (22.3%) compared to that in the 1000 s (34.0%) and 2000 s (42.6%) levitation groups, the differences were statistically insignificant ( p  > 0.05). The results suggest that acoustic levitation for less than 2000 sec does not interfere with the development of zebrafish embryos, but may affect mortality rate. Acoustic levitation could potentially be used as a non-contacting wall-less platform for characterizing and manipulating vertebrae embryos without causing major adverse effects to their development.

Effects of acoustic levitation on the development of zebrafish, Danio rerio, embryos