DRAQ7 overcomes the complications associated with autofluorescence of doxorubicin during flow cytometric analyses

Doxorubicin (DOX) is a widely used chemotherapeutic agent that induces apoptosis through DNA intercalation, topoisomerase II inhibition, and oxidative stress. However, its cardiotoxicity is a significant concern. Flow cytometry, particularly with Annexin V/propidium iodide (PI) or Annexin V/7AAD assays, is commonly used to assess DOX-induced cell death. A challenge arises from DOX's intrinsic fluorescence, which interferes with accurate detection. To address this, we successfully used DRAQ7, a far-red dye, in HeLa and other cancer cell lines. As our study demonstrates DRAQ7 eliminates auto-fluorescence interference, this approach offers precise differentiation of apoptotic cells and improving experimental and therapeutic reliability.