Tissue formation, wound healing, immune surveillance and most importantly, inhibition of cancer and tumor metastasis is of prime interest. In order to study translocation within these intriguing cells, we focus on cellular crawling motility in sperm from the nematode Caenorhabditis elegans, which is similar to the crawling motility of many other eukaryotic cells. C. elegans spermatozoa utilize a cytoskeleton composed of Major Sperm Protein (MSP) to produce the force necessary for cellular movement. A complete understanding of the mechanics of this process and how it is regulated is still lacking. In the sperm from Ascaris suum, another nematode, internal pH has been suggested as a regulator of motility. The focus of this study is to determine the role of pH in regulating motility in C. elegans sperm and to determine how the cell maintains its internal pH. Using ratiometric imaging we show that C. elegans sperm maintain a substantial pH gradient between the front and back of the cell. A mathematical model suggests how the cell maintains this gradient.