With the rapid development of Internet technology and mobile terminal equipment, online and offline blended teaching mode has developed rapidly. Based on the analysis of the problems existing in the current higher vocational course teaching and the solutions, this article introduces the exploration and practice of blended teaching in the course of Microbiology online and offline, including online teaching platform selection, teaching content of the four aspects of reconstruction, the design and implementation of teaching, teaching evaluation and assessment. At the end, The effect and reflection of the online and offline blended teaching mode was discussed. The online and offline blended teaching mode adopts fragmented teaching resources, breaks through the limitation of teaching time and space, improves the students’ self-learning ability, and improves the teaching quality of Microbiology course.