Objective In this paper, a fast, stable and sensitive model of zebrafish osteoporosis (OP) was established by comparing different methods of establishing common zebrafish osteoporosis models. Methods The OP model was induced by iron overload and prednisolone (Pred). The bone formation and animal mortality were observed. Ammonium ferric citrate (FAC) was used as the model drug in iron overload induction method. The Pred induction method was Pred-3 dpf (day post fertilization) method, the modeling time was 3 ~ 9 dpf; Pred- 5 dpf method, the modeling time was 5 ~ 10 dpf; Pred withdrawal method, 3 dpf began to make the model, and 7 ~ 9 dpf was removed every day. Results The significant effect of FAC on bone formation was not observed at the dose of 500 μg/mL. Compared with the control group, the number of bone and the length of the first vertebra in the model group induced by Pred-3dpf decreased significantly (P < 0.01, P < 0.05), but the mortality of zebrafish was higher. There was no significant difference between Pred-5 dpf method (P > 0.05) Pred withdrawal method model group were significantly reduced (P < 0.01), and no death. AC, CA and AL all had anti-OP effect, and CA had more sensitive and stable anti-OP effect. The optimization results of dyeing parameters of alizarin red (ARS) showed that the dyeing concentration was 0.02%, and the washing conditions of 0.5% KOH and glycerol for 2 hours were the best. The results of chemical staining showed that calcein was more sensitive to the staining of bone nodes and ARS was more sensitive to the staining of the first vertebra. Conclusions The zebrafish OP model induced by Pred withdrawal method is more rapid, stable and sensitive, and it is a stable and reliable model for OP study.