High-Precision Pneumatic Induction of Traumatic Brain Injury in Larval Zebrafish

Zebrafish have recently emerged as a cost-effective vertebrate model for Traumatic brain injury (TBI) research. However, current injury paradigms, particularly the weight-drop method, suffer from poor reproducibility due to sensitivity to small variations in materials and setup. Here, we introduce a high-precision pneumatic piston system, the Zebrafish Pneumatic Injury Device (ZePID), which delivers controlled, repeatable pressure pulses to induce TBI in larval zebrafish. ZePID achieves 97% accuracy in applied force and substantially reduces variability compared with weight-drop approaches. Using 6-days-post-fertilization (dpf) larvae, we quantified injury severity by tracking seizure-like locomotor behaviors. Larvae exposed to 150 psi exhibited a significant increase in maximum swimming speed relative to uninjured controls, as well as increased total distance traveled, consistent with TBI-associated hyperactivity. Overall, ZePID provides a standardized, compact, and high-efficiency method for inducing TBI in larval zebrafish.