Janus particles are asymmetric structures containing two different chemical compositions in the same particle. The amphiphilic Janus particles are two different chemical groups containing hydrophilic groups and hydrophobic groups on the opposite side, which is a new type of nanomaterial. In this paper, the amphiphilic Janus particles prepared by template method and their application fields are reviewed, including hard template method, soft template method, sacrificial template method and template-free method. The research progress of amphiphilic Janus particles in surfactants, dental adhesives, microcapsules, functional coatings, catalysts and biosensors are reviewed.