Commentary on a Classic JHC Article on the Development of Highly Sensitive Fluorochrome-labeled Tyramides for Immunocytochemistry

Abstract

This commentary reflects on the significance and impact of the highly cited companion article that was published in the Journal of Histochemistry and Cytochemistry in 1997 (Gijlswijk RPM et al. Fluorochrome-labeled Tyramides: Use in Immunocytochemistry and Fluorescence In Situ Hybridization. Journal of Histochemistry & Cytochemistry. 1997;45(3):375-382):

Sensitivity is the key to unlocking biological mysteries. The development of tyramide signal amplification (TSA) techniques in the 1990s dramatically changed the scientific landscape and opened up new avenues of experimental investigation to explore the visualization of antigens and nucleic acids in immunohistochemical (IHC), immunocytochemical (ICC), and in situ hybridization procedures. Based on a presentation at the 47th Annual Meeting of the Histochemical Society, the sponsoring organization of the Journal of Histochemistry and Cytochemistry (JHC), van Gijlswijk et al. subsequently published a groundbreaking article in JHC on the use of fluorochrome-labeled tyramides in ICC and in situ hybridization applications. ¹ Building on prior work from the Anton Raap laboratory and others, this important article described the use of fluorescent tyramides for direct detection of antigens and nucleic acids rather than indirect fluorescence detection of hapten-labeled tyramides. This advance simplified the TSA protocol, reduced background signal introduced by the hapten detection step, and facilitated the development of versatile multi-label fluorescence detection procedures.

Since this highly cited article was originally published 25 years ago, the importance of TSA as a highly sensitive and practical detection technique has been emphatically demonstrated in numerous applications. As highlighted in the original article, the remarkable sensitivity of the TSA technique permitted lower probe and antibody concentrations to be used than in conventional detection procedures resulting in decreased non-specific background, reagent cost savings, lower antigen/nucleic acid detection limits, and multi-label procedures for fluorescence in situ hybridization (FISH) and ICC/IHC detection. The advantages of fluorescence-based TSA procedures had immediate impact in both research and clinical applications and opened up entirely new avenues of investigation based on sensitive, spatially discrete detection of antigens and nucleic acids.

Fluorochrome-labeled tyramides helped usher in the modern era of multiplex antigen detection, combined ICC/IHC/FISH procedures, flow cytometric applications, combined TSA and quantum dot techniques, and highly sensitive electron microscopic protocols. Importantly, they remain highly valuable in the modern cell biology, anatomy, and experimental pathology laboratory “tool box.” This article is one of the most significant early TSA-related manuscripts and remains must reading for a new generation of dedicated scientists investigating the expression, localization, and spatial relationships of specific antigens and nucleic acids.