Significance: Endothelial c-Myc plays a critical role in redox homeostasis, with its deletion potentially exacerbating oxidative stress, particularly during chemotherapy. Aim: To assess the metabolic impact of endothelial c-Myc knockout (KO) and doxorubicin (DOXO) treatment on kidneys, hearts, and livers using 3D optical cryo-imaging. Approach: Redox ratios (NADH/FAD) were analyzed in control (CT) and KO mice treated with DOXO or saline to evaluate mitochondrial function and oxidative states. Results: KO tissues showed significant reductions in redox ratios, indicating an oxidized state, with kidneys exhibiting up to a 51.42% decrease. DOXO treatment further exacerbated oxidative stress in KO tissues, while CT groups demonstrated protective effects. Conclusions: Endothelial c-Myc is crucial for redox balance and protection against chemotherapy-induced oxidative damage, offering insights for targeted therapeutic strategies.