Environmental stressors induce specific physiological responses that can be measured in the blood, notably by morphological changes in lymphocytes. Tobacco being the best-known stress in terms of its impact on health, we studied the physiological properties of peripheral blood lymphocytes in a population of 33 healthy non-smokers and smokers. Proteasome amount, mitochondria energy levels, changes in membrane properties and cell and nuclear size were analyzed to obtain 28 parameters from two fluorescence-based techniques: flow cytometry and cell imaging. The results showed that none of the parameters alone identified gender and smoking status, but that statistical analysis of these parameters, whether or not combined with a third set of data, hematological data, can. Statistical analysis of selected parameters clearly discriminates between male and female samples, as well as smokers and non-smokers. Effects of tobacco smoke pollutants are more pronounced in female smokers than in other groups.