The morphology, the motility, and the biochemical structure of the spermatozoon have often been correlated with the outcome ofin vitrofertilization and have been shown to be the sole parameters of the semen analysis in predicting the success of intracytoplasmic sperm injection and intracytoplasmic morphologically selected sperm injection. In this context, digital holography has demonstrated to be an attractive technique to perform a label-free, noninvasive, and high-resolution technique for characterization of live spermatozoa. The aim of this chapter is to summarize the recent achievements of digital holography in order to show its high potentiality as an efficient method for healthy and fertile sperm cell selection, without injuring the specimen and to explore new possible applications of digital holography in this field.