Introduction: Dengue virus is one of the major infectious human pathogens worldwide. Currently, no antiviral drug has become available against the dengue virus induced diseases since little is known regarding how dengue virus interacts with host cells. Dengue virus-polymersome hybrid nanovesicles are powerful tools to explain the dynamics of host cell-virus interaction and tracking. Methods: Dengue virus-polymersome hybrid nanovesicles were prepared by dialysis method. We showed that DiD and BODIPY FL C5-Ceramide encapsulated dengue virus-polymersome hybrid nanovesicles triggered the formation of red and green fluorescence by using real-time fluorescence microscopy. The internalization to escape endosomal entrapment was determined by confocal laser scanning microscopy. Results: Fluorescence images of DiD and BODIPY FL C5-Ceramide encapsulated dengue virus-polymersome hybrid nanovesicles illustrated at different time points. For real-time virus tracking study, it is desirable to acquire images to obtain adequately high resolution to monitor the dynamics of host cell-virus interaction and tracking in living cells. Conclusion: This study exploited a virus-nanovesicles tracking technology to approach whether dengue virus interacts with autophagy. Therefore, we demonstrated that dengue virus-polymersome hybrid nanovesicles will be utilized for virus tracking studies to examine the mechanisms of viral infections.