Environmental protection and food safety are closely related to the healthy development of human society; there is an urgent need for relevant analytical methods to determine environmental pollutants and harmful substances in food. Molecular imprinting-based ratiometric fluorescence (MI-RFL) sensors, constructed by combining molecular imprinting recognition and ratiometric fluorescence detection, possess remarkable advantages such as high selectivity, anti-interference ability, high sensitivity, non-destruction and convenience, and have attracted increasing interest in the field of analytical determination. Herein, recent advances in MI-RFL sensors for environmental and food analysis are reviewed, aiming at new construction strategies and representative determination applications. Firstly, fluorescence sources and possible sensing principles are briefly outlined. Secondly, new imprinting techniques and dual/ternary-emission fluorescence types that improve sensing performances are highlighted. Thirdly, typical analytical applications of MI-RFL sensors in environmental and food samples are summarized. Lastly, the challenges and perspectives of the MI-RFL sensors are proposed, focusing on improving sensitivity/visualization and extending applications.