Objective: We attempt to explore the effect of microlecture combined with flipped classroom on the teaching of occupational safety protection for probationer nurses of operating room to figure out an effective teaching method to improve probationer nurses’ occupational safety protection. Methods: We grouped probationer nurses who entered the department from March 2018 to March 2019. The first to fifth crews were grouped into the control group (61 nurses), and the sixth to tenth crews were grouped into the observation group (63 nurses). Theoretical assessment model: we tested the nurses using the same test paper at the first day they came to the department and the day before they left the department. Theoretical training model: at the end of the first week of practicing, the teaching group leader gave courses on occupational safety protection to the probationer nurses practicing in the operating room. The teaching method for control group was cramming intensive teaching mode while that for observation group was microlecture combined with flipped classroom in which problem-based group discussion was adopted. Two groups’ satisfaction with the course design was investigated after class. At the fourth week prior to the end of the practicing, we assessed two groups’ skill at conducting seven-step hand-washing method and dealing with sharp instrument injury and counted the occurrence of sharp instrument injury in probationer nurses. Results: Before training, there was no significant difference in the theoretical assessment results between control group and observation group (P>0.05). After training, observation group had better results in theoretical assessment and skill operation than control group with a significant difference (P<0.05). The occurrence of sharp instrument injury of the observation group was 7.9% (5/63) while that of the control group was 21.3% (13/61). There was a significant difference (P=0.035). What’s more, there was a significant difference in the satisfaction with course design between control group and observation group (χ2=51.697, P=0.000). Conclusions: Microlecture in combination with flipped classroom is able to promote internalization and absorption of theoretical knowledge of occupational safety protection so as to improve probationer nurses’ occupational safety protection knowledge and clinical skills. The teaching mode can also reduce the incidence of sharp instrument injury in probationer nurses and thus relieve probationer nurses’ fear of occupational injuries during clinical practice. As a result, the probationer nurses are more active in the theoretical training. Therefore, microlecture combined with flipped classroom is effective on teaching the student nurses of the operating room the occupational safety protection knowledge.