Fluorescence polarization imaging can obtain the orientation and structural features of the sample by measuring the polarization parameters of fluorescent molecules, owing to which it is widely used in biology research. Moreover, it is often combined with other fluorescence microscopy techniques to analyze the arrangement of cellular structures, the real-time dynamic orientation of biomolecules, and the organization of tissues. This approach enables research on cellular physiological processes, the effects of drugs on cells, and the detection of abnormal structures. In this study, we focus on the applications of fluorescence polarization imaging in biology research, including the analysis and comparison of different methods that utilize polarization information to obtain orientation structures of different samples. Furthermore, we provide insights into future development trends in this field.