ABSTRACT
Immunohistochemical (IHC) and in situ hybridization (ISH) detection in human tissue sections have traditionally been performed with chromogenic substrates due to significant autofluorescence in many human tissues and the relatively poor sensitivity and photostability of conventional fluorescence detection procedures. Recent advances in enzyme-based amplification methods, novel fluorophores, and improved detection protocols have dramatically increased the utility of fluorescence localization procedures in human tissue sections. This chapter provides detailed protocols for enzyme-enhanced immunohistochemical and ISH detection and discusses practical solutions to common problems encountered using these sensitive methods.
