Due to the specific recognition performance, imprinted polymers have been widely investigated and applied in the field of separation and detection. Based on the introduction of the imprinting principles, the classification of imprinted polymers (bulk imprinting, surface imprinting, and epitope imprinting) are summarized according to their structure first. Secondly, the preparation methods of imprinted polymers are summarized in detail, including traditional thermal polymerization, novel radiation polymerization, and green polymerization. Then, the practical applications of imprinted polymers for the selective recognition of different substrates, such as metal ions, organic molecules, and biological macromolecules, are systematically summarized. Finally, the existing problems in its preparation and application are summarized, and its prospects have been prospected.