Abstract:Sperm cells of rice ( Oryza sativa L. ) were isolated from mature pollen grains near anthesis using osmotic shock with sucrose solution. They were separated from pollen contaminants by a Percoll gradient centrifugation. Isolated sperm cells were viable as tested with the fluorochromatic reaction. Cytological observations revealed that the two sperm cells just released from a pollen grain were linked together and each had a long tail-like extension. The isolated sperm cells were spherical with irregular surface and most of them separated, but some still remained in pairs that connected by finger-like structure. As observed with TEM, the isolated sperm cell was surrounded by a single plasma membrane and cell wall did not exist. Sperm nucleus was large and euchromatic.