This work described the effect of furfural, a product resulting from the lignocellulosic material pretreatment, on Saccharomyces carlsbergensis growth and ethanol production. Flow cytometry was used to evaluate the yeast membrane potential, membrane integrity, reactive oxygen species production and lipid content. Above 0.3 g/L of furfural, a progressive decrease in the maximal specific growth rate was observed, reaching 53% of the value obtained in the absence of toxic when the cells were grown in the presence of 4 g/L of furfural. In general, the yeast biomass concentration and yield were less affected by the furfural presence than the specific growth rate, and a maximum reduction of 25% was observed for the assay at 4 g/L. The ethanol production was even less affected by the furfural presence than the yeast growth. At 4 g/L of furfural, the maximum ethanol concentration was reduced by only 10% relatively to the maximum ethanol concentration observed in the absence of toxic. At 5 g/L of furfural, the yeast cells were barely able to keep metabolic functions and produced a final ethanol concentration of 0.87 g/L although growth was undetectable. S. carlsbergensis membrane potential was affected by the furfural presence, concomitantly with the ethanol production. However, at 4 g/L, most of the yeast cells (90%) displayed the cytoplasmic membrane depolarized. The proportion of cells with increasing reactive oxygen species (ROS) production levels increased for the experiments at 0-4 g/L. For the experiment at 4.5 g/L of furfural, ROS production was observed for only 11% of the yeast cells. The yeast lipid content was also severely affected by the furfural presence. Both polar and neutral lipids decreased in the presence of furfural, and this reduction was more notorious during the stationary phase.