Objective: Freeze and thaw have some detrimental effects on sperms. Molecular changes on the sperm surface can impact on fertility rate. We tested wheat germ agglutinin (WGA) peanut agglutinin (PNA) and Dolichos biflorus (DBA) to detect surface glycoconjugates before and after freezing Materials and Methods: For this purpose 45 healthy semen samples were frozen and thawed and sperm smears were prepared before and after freezing and thawing. The smears were stained with the lectins and also with acridin orange. The smears were studied by fluorescents microscopy and the intensities of the reactions to lectins were measured by image analyses software. Results: The results indicated that the reactions of 46.67% 34.09% and 73.34% of the specimens modified to PNA WGA and DBA lectins respectively after freezing and thawing. The cryopreservation caused both increase and decrease of the intensities of the reactions. It means that there are various mechanisms that impact on the carbohydrate contents of the sperm surface. Conclusion: We conclude that cryopreservation affected the surface glycoconjugates at least in a subset of spermatozoa. These results might improve future application of sperm banking techniques.