Exosomes are tiny vesicles secreted by cells that carry proteins, nucleic acids, lipids, and cellular messenger molecules, participating in intercellular communication and possessing potential applications in cancer diagnosis, drug delivery, and other fields. However, due to their small size and environmental sensitivity, the preservation of exosomes presents challenges. This study investigated the effects of temperature and storage time on the stability of urinary exosomes, as well as the impact of pre-urination diet on exosomes. A specialized slide was designed to effectively immobilize and display exosomes, providing a cost-effective solution. Results showed that exosome stability was optimal when frozen at -20°C. Further experiments demonstrated that consuming large amounts of water before urination resulted in higher exosome concentrations, while high-sugar beverages increased the size of nanoparticles, resulting in a decrease in the number of exosomes encapsulated within the exosome envelope. This study provides new insights and methods for the preservation and characterization of urinary exosomes, both in basic and applied research on exosomes.