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Glucose oxidase-dependent deposition of fluorochromized tyramide (FT-GO) for sensitive multiplex immunofluorescence and in situ hybridization labeling

Highlighted Citation

Tyramide Signal Amplification (TSA) uses peroxidase (POD) catalysis to produce high-intensity labeling of biomolecular targets and enables the detection of low-abundance targets in immunocytochemistry (ICC), immunohistochemistry (IHC), and in situ hybridization (FISH) applications. Specifically, POD reacts with hydrogen peroxide (H₂O₂) to catalyze production of reactive tyramide radical intermediates, which covalently bind to electron-rich molecules such as tyrosine, phenylalanine, and tryptophan amino acids near the site of catalysis. The ability of POD to catalyze the deposition of many fluorochromes at the labeling site results in signal amplification and a significant increase in sensitivity compared to traditional immunofluorescence (IF). Multiplex TSA-based imaging has emerged as a central tool in spatial biology studies, with several comprehensive kits commercially available along with various stand-alone TSA components for customized and cost-effective panel design. While multiplex TSA staining has become an indispensable tool, further refinement could make the technique more sensitive and accessible.

In a recent Scientific Reports article, Yamauchi et al. eloquently showed the utility of a novel multiplex TSA method using a custom panel of Biotium’s CF® Dye Tyramides. A limitation of standard TSA is that...