Abstract: Objective Based on the present teaching situation of physiology experimental course of higher vocational nursing specialty in our college, we discuss the implementation method and application effect of flipped classroom teaching mode based on micro-lecture in physiology experimental teaching, in order to provide reference for the promotion of this teaching mode in physiology experimental teaching. Methods The study object selected class 1 and 2 of three years higher vocational nursing specialty in our college, and class 1 was the control group (n=40)， and class 2 was the experimental group (n=40). The traditional teaching mode was used in the control group, and the experimental group used the teaching mode of flipped classroom on basis of micro-lecture. The flipped teaching based on micro-lecture mainly included four steps, namely, teachers' preparation before class, students' independent learning before class, classroom implementation and consolidation after class. After the end of the course, the teaching effects of two groups of students were evaluated in experimental theory, experimental skill and questionnaire investigation. We used software SPSS 19.0 in the statistics of data. T-test was used in comparing between groups, and we used χ² test in the comparison of count data. Results The experimental group's average score of experimental theory test [(84.85±7.43) points] was more than control group's [(80.47±7.69) points], and the experimental group's average score of experimental skill test [(83.93±7.48) points] was also more than control group's [(78.78±6.44) points], and the differences were statistically significant (all P<0.05). The results of the questionnaire survey showed that the experimental group students' recognitions of the teaching mode were better than that of the control group in learning interest, self-learning ability, awareness of teamwork and ability to analyze and solve problems, and the differences were statistically significant (all P<0.05). Conclusion The flipped classroom teaching mode based on micro-lecture can enhance students' mastery of experimental skill and experimental theory, help to improve students' interest in learning, develop students' self-learning ability, enhance students' awareness of teamwork and improve students' ability to analyze and solve problems. Therefore, this teaching mode deserves further study and promotion in the physiology experimental teaching.