AIM:This study aimed to investigate the value of DNA ploidy and galectins-3 immunostain in the preoperative evaluation of thyroid epithelial lesions. MATERIAL AND METHODS:Sixty patients presenting with thyroid enlargement were included in this study and referred by clinicians for FNA. Routine cytological evaluation was done on PAP stained slides according to the WHO criteria and at least three slides were prepared for routine cytological examinations. The nuclear DNA analysis was performed at the Pathology Department, National Research Center using the Leica Qwin 500 Image Analyzer (LEICA Imaging Systems Ltd, Cambridge, England). Galectin-3 expression was investigated in all tissues using streptavidin-biotin technique. RESULTS:Conventional Fine needle aspiration cytology (FNAC) of 60 cases could diagnose malignancy with a sensitivity of 60%, negative predictive value (NPV) 71.4%, and overall diagnostic accuracy of 80%. The aneuploidy was significantly associated with malignancy, with sensitivity 90.9%, specificity 83.3% and accuracy 88.3%. On using galectin-3 immunocytochemichal stain on cell blocks prepared from FNA the values were improved, sensitivity 93.3% specificity 86.7% and overall accuracy 90% and it was noticed that galectin-3 over expression was significantly associated with malignancy. CONCLUSIONS:From the results of this study we can consider that DNA ploidy and Galectin-3 could refine the FNA results and increase its sensitivity as a screening test from sensitivity(60%) to reach sensitivity (93.3%), thus decreasing the false negative cases. From this study, it is concluded that the application of ancillary techniques as galectin-3 immunocytochemical markers may become a reliable indicator for surgical intervention, DNA ploidy measurements on the other hand may be of value in galectin-3 negative cases to determine the behavior of the lesion in such cases & refine the preoperative assessment by out ruling false negative cases.