Abstract: Microbiology experiment technology is an important part of the teaching content of Microbiology. The traditional teaching mode makes the repetitions of microbiology experiment teaching and key experimental were limited. And it is difficult for students to grasp the essentials of some experimental operation. At present, multimedia teaching is an important supplementary means in microbiology teaching of university. By the researching and application of multimedia courseware in microbiology experimental teaching, the experimental teaching of microbiology become intuitive and substantial; teaching resources was saved, the problem of the lack of experimental facilities and teaching difficulties were solved, the teaching effect of microbiology experiment was greatly improved.