Abstract: In poplar, previous studies on the molecular mechanisms of male gamete production used whole anthers or even whole inflorescence, therefore they cannot provide celltype specific expression information. This study establishes a system separating poplar microspore and pollen cells at different developmental stages by using the frozen section and laser microdissection technologies. Further extraction of poplar RNA was conducted and our results showed that the integrity and purity had achieved the requirements of linear application. This study can facilitate our research towards more precise molecular mechanisms of male gamete production and even a more comprehensive understanding of male gamete production in poplar.