Abstract:A technique named “two-step osmotic shock” was developed for the isolation of large number of generative cells from pollen grains in Vicia faba and Zephyranthes candida. The procedure included: 1. Pollen hydration: mature pollen grains were incubated in adequate sucrose solution for 30-50 min. 2. Osmotic shock, when the pollen grains swelled and showed the tendency of germination, a sudden osmotic shock by adding equal volume of water made them bursting. The contents carrying generative cells spurted from the germ pores. By subsequent filtration and centrifugation, purified generative cell population was obtained. FDA test proved that the isolated and purified generative cells were viable.