Graphical Abstract (IMAGE) Shanghai Jiao Tong University Journal Center Caption The discovery of G protein-coupled receptor (GPCR) heteromers has revealed a new level of complexity in their functional characteristics. This review explores a variety of methodologies and live cell imaging technologies for the investigation of physical interactions in GPCR heteromers and their functional alterations. Live cell imaging technologies such as fluorescent protein-based biosensors promise a deeper understanding of GPCR heteromers, urging a reassessment of their physiological importance and pharmacological relevance. Credit All authors (Hyunbin Kim, Huimin Lee & Jihye Seong) Usage Restrictions Credit must be given to the creator. Only noncommercial uses of the work are permitted. No derivatives or adaptations of the work are permitted. License CC BY-NC-ND Disclaimer: AAAS and EurekAlert! are not responsible for the accuracy of news releases posted to EurekAlert! by contributing institutions or for the use of any information through the EurekAlert system.