Fibroblast growth factor 1 (FGF1) has been shown to maintain proliferation and self-renewal capacities of neural stem/progenitor cells (NSPCs) in vitro. We have previously identified FGF1B as the major transcript of FGF1 gene expressed exclusively in brain areas that are known to be abundant for NSPCs in vivo. The 540-bp (-540 to +31) sequence upstream of the 1B transcription start site (F1B) is sufficient to drive the expression of a heterologous luciferase reporter in cultured cells. In this study, we report a direct genetic and functional approach to isolate F1B(+) NSPCs using green fluorescent protein (GFP) reporter gene under the control of human F1B promoter. The F1B-GFP reporter could facilitate the isolation of NSPCs with self-renewal and multipotent capacities from human glioblastoma tissues, developing or adult mouse brains by fluorescence-activated cell sorting. Future work elucidating the mechanisms that control FGF1B expression will help to identify new NSPC-related genes.