In-depth investigations are needed on methods of semen collection and analysis in endangered nonhuman primates. Due to lack of resources, zoos need the assistance and support of universities and primate centers to perform these investigations. Several collection methods are described. Penile and rectal probe electrical stimulation methods for semen collection are compared. Rectal probe methods have been historically used in zoos; however, more representative samples may be acquired from primates indoctrinated to penile stimulation methods. Semen volume, concentration, pH, along with sperm morphology and motility are the commonly analyzed parameters for primates. However, with additional application to primate semen of motion analysis, viability tests, acrosomal analysis, hamster cell penetration tests, and sperm penetration tests, more accurate assessment of seminal capability may become routine. The examiner must be aware of the social and health situations of the primate under analysis, since any or all of these parameters can be adversely affected. The lion-tailed macaque (Macaca silenus) is used as an example for application of these analyses. The application of these analytical techniques to primate populations should allow better assessment about a primate's reproductive potential and the potential use of these semen samples for artificial techniques.