This study was conducted to investigate protocol standardization for cryopreservation spermatozoa of the bar-tailed flathead Platycephalus indicus. The suitability of the cryoprotectants, dimethyl sulphoxide (DMSO), glycerol and methanol were tested against three freezing rates and three thawing temperatures. DMSO and glycerol gave significantly higher motile index and survival rates than methanol. Among the freezing rates, freezing at a height of 2 cm above LN2 surface for 10 min-1 gave higher motile index and survival rates. In terms of best thawing temperature, 20?C obtained the highest motility.