Laser Raman spectroscopy (LTRS) is used to monitor the Raman spectroscopic changes of Cryptococcus humicolus cells and mitochondria stressed by aluminum (Al). The results show that the characteristic Raman peaks of C. humicolus cells and mitochondria are significantly decreased with the increase of aluminum concentration and treatment time, the intensity of Raman peaks associated to nucleic acid, protein and lipid respectively exhibits significant decreasing tendency with the extension of treatment time, indicating that the content of nucleic acid, protein and lipid in the cells and mitochondria is reduced during the apoptosis process. The intensity of characteristic peaks related to cytochrome c (Cyt c) in mitochondria significantly decreases with the increase of aluminum concentration and treatment time, suggesting that the yeast mitochondrial membrane rupture leads to Cyt c release into outer of mitochondria. With the increase of aluminum concentration and treatment time, the respiratory peak is observed to decrease gradually, indicating that the mitochondrial activity becomes weaker, energy metabolism has been hampered. LTRS is used for real-time monitoring of the dynamic process of cell apoptosis and physiological and biochemical changes of in vitro mitochondria under aluminum stress, and revealing the release behavior of Cyt c during the apoptosis process of C. humicolus cells induced by aluminum. The results help understand the biological mechanism of aluminum toxicity in acidic soils on organisms.