Automatic High-Throughput Injection System for Zebrafish Larvae Based on Precise Positioning of Injection Target

Zebrafish microinjection is widely used in vascular biology, neurology, and other research areas. The microinjection of zebrafish larvae is a complicated 3D task because zebrafish larvae are independent individuals with complex body structures compared with biological cells. In this study, we propose an automatic high-throughput injection system for zebrafish larvae under an optical inverted microscope. The proposed system combines the fixing device design, object detection, visual positioning, and 3D injection path planning to improve the survival rate of zebrafish after injection. Experimental results demonstrated the capability and efficiency of the proposed system, which achieved a success rate of 95.0% and a survival rate of 98.7% for zebrafish injection. The system can be applied to various biomedical and biochemical experiments.
Note to Practitioners
—Zebrafish larvae have long been an important model organism in neuroscience, biomedicine, and drug discovery. In various methods for external substance transportation into the zebrafish, microinjection is more efficient but also more difficult. At present, high-throughput injection of zebrafish larvae is mainly based on traditional manual operations. The injection speed and success rate of manual injection gradually decrease with the increase in zebrafish numbers. In this study, an automatic injection system is designed and implemented for high-throughput zebrafish larvae injection. We first use a simply designed mold to fix batch zebrafish larvae. Then we propose a coarse-fine two-step positioning method and perform a 3D injection path planning to achieve precise microinjection. The proposed injection system frees the operators from the repetitive and tedious injection work, providing technical support for the life science experiment.