At group cells level and a single yeast level, laser tweezers Raman spectroscopy was used to monitor the dynamic of the intracellular biological macromolecular in realtime during the apoptosis process of yeast cells stressed with high concentrations of ethanol. The results show that the intensities of Raman peaks of nucleic acids (721, 1 083, 1 301 cm－1), proteins (721, 858, 1 001, 1 301, 1 445, 1 608, 1 657 cm－1) and lipids (1 083, 1 301, 1 445 cm－1) decreased significantly along with the extension of treatment time, indicates that the contents of nucleic acids, proteins and lipids are reduced gradually while the yeast cells are undergoing apoptosis induced by high concentrations of ethanol. However, the results of single factor analysis and repeated measures analysis show that the changes of the intensities of Raman peaks between the group cells and the single cells are difference significantly, implies that the heterogeneities of the single cells are covered by the average spectroscopy of the population cells. The results display that laser tweezer Raman spectroscopy can be used to directly and truthfully detect the kinetic apoptosis process under high concentrations of ethanol stress at the single cell level and probe cellular heterogeneity.