Abstract: Laboratory animals play an important and fundamental role in life sciences, and their great contribution comes at the cost of various physical and mental injuries, pains, and even deaths. As a key model organism, zebrafish has become one of the most widely used laboratory animals in recent years. An important issue for researchers is how to protect zebrafish welfare while ensuring the reliability of experimental results. Here, we systematically review key welfare concerns in zebrafish research, including pain and pressure perception, water environment, feeding and breeding, experimental operation, transportation, and euthanasia. This review provides theoretical support and practical guidance for improving the life quality of zebrafish and promoting the standardization of laboratory fish.