Fluorescence in situ hybridization method for co-localization of mRNA and GFP

Co-localization studies using green fluorescent (GFP) and fluorescence immu nohistochemistry have become commonplace. However, co-localization studies using GFP and nRNA in situ hybridization are rare, in large part because ty pical in situ hybridization reaction conditions often lead to the loss of G FP fluorescence. Here, we describe a new fluorescence mRNA in situ hybridiz ation protocol using cRNA riboprobes that leaves GFP fluorescence intact. T his protocol is based on a urea-based hybridization buffer and the Tyramide Signal Amplification (TM) system. This protocol should provide researchers engaged in the use of GFP with a solid starting point for adapting their o wn in situ hybridization protocols.