This article introduced the basal components and structures of Ussing chamber and its applications in the field of gastrointestinal barrier function and nutrient transport research. During the process of applying Ussing chamber system, some problems had been found and the solutions had been proposed. It was suggested that in vitro environment needed to be accorded with gastrointestinal physiological characteristics for accurate experimental results and meaningful physiological simulation, simultaneously, material cost was decreased by improvement of methods.［Chinese Journal of Animal Nutrition, 2010，22（3）:511-518］