While Annexin V staining is a powerful tool, it has limitations. It primarily detects early apoptosis and may not distinguish between cells undergoing apoptosis or other forms of programmed cell death, such as necrosis or autophagy. Additionally, cells in advanced stages of apoptosis may not retain phosphatidylserine on their surface, leading to false negatives. Therefore, it is often used in conjunction with other assays for a comprehensive analysis.
