A sperm computer analysis and immunocytochemistry of separated seminal plasma proteins during low temperature storage of ram spermatozoa were carried out. It was found that low molecular weight seminal plasma proteins reliably protect the sperm motility (24.91% against 4.66% of progressively motile sperm at p<0.001). It was also observed that the velocity of sperm cells from sample 11 was better than in sample 3 (respectively 33.23% against 1.9% of the high motile sperm at p<0.001). Immunocytochemical analyses visualized that some of the seminal plasma proteins (SPPs) attached to the plasma membrane in the apical region of the head, midpiece and sperm tail. This gives us a good reason to suggest that these attached proteins may affect cell vitality. In conclusion, seminal plasma proteins with low molecular weight selected chromatographically contained molecular factors that could have protective effects on the ram sperm during in vitro storage at low temperatures.