The purpose of this study was determined suitable equilibration method and collection of spetmatophore to formulate technical protocol of spermatophores cryopreservation of black tiger shrimp Penaeus monodon in liquid nitrogen. The two equilibration methods (one step, two steps) and three equilibration time (25,30, 40 min), three collection of spermatophore have been tested. The results show that tiger shrimp spermatophore preserved in calcium-free saline solution, dimethyl sulfoxide (DMSO) 5 percent, used two steps equilibration method, equilibration time 25 min, collection of spermatophore by stripping, membrane separation, mix well for survival rate highest 32.53 +/- 1.29 percent (p 0.05).