The article discusses the standardization of sample collection, isolation, and analysis methods for extracellular vesicles (EV) and extracellular RNA (exRNA) in research. It highlights the growing interest in EV as potential biomarkers and therapeutic targets, particularly in oncology. However, the field faces challenges in standardizing techniques, understanding EV subtypes, and ensuring reproducibility of results. A special workshop of the International Society for Extracellular Vesicles (ISEV) in 2012 aimed to address these issues by discussing best practices for EV isolation, RNA purification, and therapeutic applications. The conclusions from the workshop, along with a review of existing literature, emphasize the need for standardized protocols, appropriate controls, and consistent techniques to facilitate comparison of results across studies. The article also addresses the importance of defining biofluid-specific norms, considering factors such as age, gender, and health status, and ensuring ethical and legal compliance in sample collection and processing. It provides detailed recommendations for sample collection, handling, and processing in various biofluids, including blood, saliva, urine, nasal secretions, breast milk, and cerebrospinal fluid (CSF). The text also discusses the challenges of isolating EV from different sources, the impact of anticoagulants on EV isolation, and the importance of controlling for variables such as fasting status and time of sample collection. The article concludes with a call for continued research and standardization to improve the reliability and utility of EV research in clinical and diagnostic applications.The article discusses the standardization of sample collection, isolation, and analysis methods for extracellular vesicles (EV) and extracellular RNA (exRNA) in research. It highlights the growing interest in EV as potential biomarkers and therapeutic targets, particularly in oncology. However, the field faces challenges in standardizing techniques, understanding EV subtypes, and ensuring reproducibility of results. A special workshop of the International Society for Extracellular Vesicles (ISEV) in 2012 aimed to address these issues by discussing best practices for EV isolation, RNA purification, and therapeutic applications. The conclusions from the workshop, along with a review of existing literature, emphasize the need for standardized protocols, appropriate controls, and consistent techniques to facilitate comparison of results across studies. The article also addresses the importance of defining biofluid-specific norms, considering factors such as age, gender, and health status, and ensuring ethical and legal compliance in sample collection and processing. It provides detailed recommendations for sample collection, handling, and processing in various biofluids, including blood, saliva, urine, nasal secretions, breast milk, and cerebrospinal fluid (CSF). The text also discusses the challenges of isolating EV from different sources, the impact of anticoagulants on EV isolation, and the importance of controlling for variables such as fasting status and time of sample collection. The article concludes with a call for continued research and standardization to improve the reliability and utility of EV research in clinical and diagnostic applications.