Introduction: The acrosome is a sac shaped organelle contaning lysosomal enzyme that is located on the anterior of sperm head. During the acrosome reaction its content is released in the vicinity of the egg. The acrosome reacted sperm then penetrates through the egg barriers. There are several methods for evaluating the acrosomal status; some of those are complex, expensive and/or non-accurate methods. The aim of this study was comparison between three of these methods, quinacrine fluorescent and current histochemical double & triple staining methods. Material and Methods: Ten norm al ejaculate semen samples were selected according to the WHO, 1999 criteria. The motile sperm were liberated from seminal plasma using percoll preparation. The acrosome reaction was then evaluated before and after capacitation, following induction by calcium ionophore for 1 to 2 hrs using three different quinacrine, double and triple staining methods. Results: There was no significant difference between the three different staining methods (P> 0.05), while after capacitation and induction using calcium ionophore the acrosome reaction increased significantly (P< 0.01). In all experiments, the percentage of acrosome reaction detected in quinacrine was significantly higher than the other methods (P< 0.01). There was a direct correlation among different methods (P< 0.001, r=7). Conclusion: The acrosome reaction occurred in sperm following capacitation and induction by calcium ionophore. The quinacrine method is a rapid and simple method for detecting the acrosomal status of human spermatozoa.