Abstract: The germination of pollens and the isolation and collection of sperm cells were studied by using a Phalaenopsis hybrid in this paper. The results show that the nucleus of generative cell degenerated after the pollen germinated and the generative cell moved into the pollen tube. The generative cell was still in cell division after pollinated 5 days. Pollen tubes were induced in the ovary after manual pollination and the sperm cells were isolated from pollen tubes by immediately blowing up in a broken solution containing 5%–12% mannitol. Four to five pairs of sperms could be collected within 30 mins by using a micromanipulator. The isolated pairs of two sperm cells shown significant differences in sizes and fluorescent intensities，that suggested both sperms have different futures. The successful isolation of sperm cells from Phalaenopsis wouldprovide a solid basis for the studies of in vitro fertilization in orchid in the future.

Key words: Phalaenopsis hybrid, generative cell, sperm cell isolation, sperm cell collection, pollen tube blow up