Improved localization of fluorescent tyramides for fluorescence in situ hybridization using dextran sulfate and polyvinyl alcohol.

Abstract

Recently, a peroxidase-mediated amplification system has been described for immunofluorescence and fluorescence in situ hybridization studies. It is based on the deposition of hapten- or fluorochrome-labeled tyramide molecules. Although providing a significantly increased detection sensitivity compared to conventional procedures, its localization properties are inferior because of free diffusion of intermediate reaction products before they are immobilized. In enzyme cytochemistry, it is well established that improved localization of enzyme activity can be achieved through the addition of viscosity-increasing polymers to the incubation media. In this study we analyzed the effect of different polymers on the localization sharpness and sensitivity of the tyramide-peroxidase reaction in FISH applications. Significantly improved localization of the fluorescent endproduct was observed using dextran sulfate or polyvinylalcohol (PVA) with, respectively, no or little loss of sensitivity.

Novel Application of Tyramide Signal Amplification (TSA): Ultrastructural Visualization of Double-labeled Immunofluorescent Axonal Profiles

“…1996; van Gijlswijk et al 1996van Gijlswijk et al ,1997Van Heusden et al 1997). Essentially, in the TSA technique the biotinor fluorophore-conjugated tyramide is "attracted" to the antigenic site activated by the peroxidase component of the immunolabeling complex.…”

Tyramine Amplification Technique in Routine Immunohistochemistry

“…However, in our hands the protocol described here offered the best compromise between simplicity, reliability, and beneficial increase in sensitivity (or antibody dilution). For even more increased sensitivity, additional modifications in the TAT protocol may be useful, e.g., repetition of tyramine incubation and/or prolongation of the tyramine incubation period through the use of highly viscous diluents (Merz et al 1995;van Gijlswijk et al 1996).…”

Role of the receptor for advanced glycation end products inhepatic fbrosis

“…Sections were developed using Fast Red and nuclei counterstained with hematoxylin. RAGE was additionally detected with the catalyzed signal amplification system as previously described [29,20] . For double staining, the slides were treated with a Double Staining Enhancer (Zytomed 50-056) for 30 min before application of the secondary antibody.…”