Fluorescent Tyramide
Fluorescent Tyramide - Related Videos
14:21
High Resolution Fluorescent In Situ Hybridization in Drosophila Embryos and Tissues Using Tyramide Signal Amplification
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Cited by 25 •
2017
The described RNA in situ hybridization protocol allows the detection of RNA in whole Drosophila embryos or dissected tissues. Using 96-well microtiter plates and tyramide signal amplification, transcripts can be detected at high resolution, sensitivity, and throughput, and at a relatively low cost.
04:06
Performing Double Fluorescence In Situ Hybridization to Detect Gene Expression in Mouse Brain Sections
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2025
Source: Jolly, S., et.al. Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections. J. Vis. Exp. (2016).This video demonstrates the use of double fluorescence in situ hybridization to visualize the expression of two RNA targets in mouse brain sections. The protocol involves sequential detection of RNA using probes labeled with fluorescein isothiocyanate and digoxigenin, followed by enzymatic signal...
07:37
Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals − Using Unconjugated Primary Antibodies from the Same Host Species
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Cited by 6 •
2020
Several different methods have been established for multiplex immunostaining using primary antibodies from the same host species. Here, we describe the use of microwave-mediated antibody stripping and of fluorophore-tyramide to block antibody cross-reactivity during multiplex immunostaining on formalin-fixed paraffin-embedded mouse adrenal sections.
09:15
Tyramide Signal Amplification for the Immunofluorescent Staining of ZBP1-Dependent Phosphorylation of RIPK3 and MLKL After HSV-1 Infection in Human Cells
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Cited by 1 •
2022
Tyramide signal amplification during immunofluorescent staining enables the sensitive detection of phosphorylated RIPK3 and MLKL during ZBP1-induced necroptosis after HSV-1 infection.
09:23
Combining Double Fluorescence In Situ Hybridization with Immunolabelling for Detection of the Expression of Three Genes in Mouse Brain Sections
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Cited by 15 •
2016
Localizing gene expression to specific cell types can be challenging due to the lack of specific antibodies. Here we describe a protocol for simultaneous triple detection of gene expression by combining double fluorescence RNA in situ hybridization with immunostaining.
FAQs
The amplification depends on a localized chemical sequence. Horseradish peroxidase acts on fluorescently labeled tyramide when hydrogen peroxide is present, generating reactive radicals. These radicals attach the fluorescent reagent covalently to proteins close to the detection site, producing a concentrated fluorophore deposit rather than relying on a single nearby label.
Signal strength increases because many fluorescent molecules can accumulate around one detection site. Their covalent attachment keeps the deposited label spatially close to the target, combining enhanced visibility with precise localization. This arrangement is particularly useful when the protein or nucleic acid target occurs at low abundance and would otherwise produce a difficult-to-visualize fluorescence pattern.
Horseradish peroxidase provides the enzyme activity that converts fluorescently labeled tyramide into reactive radicals, while hydrogen peroxide enables that conversion. The resulting radicals are responsible for attachment near the detection site. Because both components participate in the activation step, they connect enzymatic activity with localized fluorescent deposition in the assay.
A target is first identified through an assay format such as fluorescence in situ hybridization or immunofluorescence, with horseradish peroxidase positioned as part of the detection system. Fluorescently labeled tyramide and hydrogen peroxide then support radical formation and local deposition. Imaging afterward reveals the strengthened fluorescence pattern around the target site.
In genetics, the approach can strengthen localization signals for genes, transcripts, and proteins. It is applicable to fluorescence in situ hybridization, immunofluorescence, and related assays performed in cells or tissues. The resulting patterns help researchers examine where molecular expression occurs, including targets whose abundance is too low for easily visible detection.
Its localized deposition allows one detection event to place fluorophores near the relevant molecular site without broadly distributing signal across the sample. That spatial precision supports sequential staining and multiplex imaging, in which researchers analyze multiple molecular signals in the same specimen. These capabilities help compare expression patterns across genes, transcripts, or proteins within cells and tissues.