Single extracellular vesicle (EV) analysis holds great promise for non-invasive cancer diagnostics, offering insights into tumor-specific biomarkers and enabling personalized treatment strategies. However, a significant challenge in the path towards clinical applications is the low abundance of tumor-derived EVs (s) in biofluids, which reduces the sensitivity, specificity, and accuracy of detection. This review emphasizes the importance of analyzing a large number of single EVs to overcome this limitation. We estimate that less than 0.1% of total EVs could be from cancer cells in a mixed sample. Additionally, the development of more efficient s isolation methods and targeted enrichment strategies, as well as high-throughput analysis techniques are crucial for improving diagnostic accuracy and advancing liquid biopsy applications in cancer care.