Abstract : The paired box gene 3 (Pax3) is expressed during pigment cell development. We tested whether the targeted allele Pax3(GFP) can be used as a reporter gene for pigment cells in the mouse. We found that the enhanced Green Fluorescent Protein (GFP) can readily be seenin every melanoblastand melanocyte in the epidermis and hair follicles of Pax3(GFP/+) heterozygotes. The GFP was detected at all differentiation stages, including melanocyte stem cells. In the dermis, Schwann cells and nestin-positive cells of the piloneural collars resembling the nestin-positive hair follicle multipotent stem cells exhibited a weaker GFP signal. Pigment cells could be purified by fluorescent activated cell sorting and grown in vitrowithout feeder cells, giving pure cultures of melanocytes.The Schwann cells and nestin-positive cells of the piloneural collars were FACS-isolated basedon their weak expression of GFP. Thus Pax3(GFP) can discriminate distinct populations of cells in the skin. Significance:? In this report, the Pax3(GFP) allele is validated as a convenient and sensitive reporter for pigment cells in the epidermis and hair follicles. Pax3(GFP) provides a FACS usable marker formelanoblasts and melanocytes from embryonic and neonatal skin and can be used to obtain pure cultures of melanocytes. © 2012 John Wiley & Sons A/S.