The comet assay is a relatively inexpensive, fast, sensitive and reliable method to detect DNA breakage upon individual cells. The most widely used cell type in this technique is lymphocyte, because they are easy to obtain and handle, and because they are continually exposed to xenobiotics that enter into the body. However, is important to consider the possibility to use other cell types for very specific purposes. Sperm cells are of special interest because they could be used as a biomonitor to risk assessment in populations occupationally exposed to xenobiotics. Besides, the fact that there are not functional DNA repair mechanisms in these cells could increase the sensitivity of the system. We present here several modifications to the comet assay methodology to evaluate DNA breakage in sperm cells with reliable results.