TSA and Styramide™ Imaging

AAT Bioquest offers a comprehensive range of tyramide signal amplification (TSA) and power Styramide™ signal amplification (PSA) reagents for ultrasensitive detection of low-abundance targets in immunohistochemistry (IHC), immunocytochemistry (ICC), and fluorescence in situ hybridization (FISH). The Styramide™ system represents a significant advancement over traditional TSA, providing enhanced signal intensity, improved photostability, and superior water solubility through our proprietary iFluor® dye technology. These products enable researchers to detect targets that would otherwise be invisible with conventional immunofluorescence methods.

Product Type	Technology	Best For	Key Advantage
iFluor® Styramide™ Conjugates	PSA	Maximum sensitivity IHC/ICC	Superior brightness & photostability
PSA Imaging Kits	PSA	Complete workflow solution	All-in-one kit with HRP secondary antibodies
iFluor® Tyramide Conjugates	TSA	Standard TSA applications	Improved over classic tyramides
mFluor™ Styramide™ Conjugates	PSA	Large Stokes shift applications	Reduced spectral overlap
Hapten Styramide™ Conjugates	PSA	Hapten-based detection	Flexible secondary detection
Classic Tyramides	TSA	Cost-effective TSA	Older technology
Buffers & Accessories	TSA/PSA	Protocol optimization	Enhanced signal & reduced background

Fig. 1

Sensitivity of power Styramide™ signal amplification (PSA) kit. HeLa cells were fixed, permeabilized, and labeled with various concentrations of rabbit anti-tubulin primary antibody as per manufacturer's recommendation. Cells were then stained with reagents in iFluor® 488 PSA Imaging Kit with goat anti-rabbit IgG, an Alexa Fluor® 488-labeled tyramide, or an Alexa Fluor® 488-labeled goat anti-rabbit IgG. Cell images were captured from each treatment under the same conditions (using a FITC filter set and analyzed with the same exposure time). Relative fluorescence signal intensity was measured and compared between different signal amplification systems.

iFluor® Styramide™ Conjugates

iFluor® Styramide™ conjugates utilize AAT Bioquest's proprietary power Styramide™ signal amplification (PSA) technology, which represents a major advancement over traditional tyramide signal amplification. The Styramide™ chemistry provides enhanced covalent deposition efficiency, resulting in significantly brighter signals with lower background compared to conventional tyramide reagents.

Key Features of PSA Technology

Up to 100× brighter signals than IHC
10-50× brighter signals than tranditional tyramide
Superior photostability for extended imaging sessions
Enhanced water solubility reduces non-specific background
Full spectral coverage from UV (350 nm) to near-IR (860 nm)
Compatible with standard HRP-based detection systems
Ideal for multiplex immunofluorescence with minimal spectral overlap

Fig. 2

Power Styramide™ signal amplification (PSA) is a highly sensitive method for detecting low-abundance targets in cells and tissues. Compared to traditional tyramide signal amplification (TSA), PSA produces ~10–50× higher fluorescence signal and can deliver >100× higher sensitivity when used with iFluor® dye-labeled Styramide™ conjugates. PSA uses HRP-catalyzed covalent fluorophore deposition for rapid, robust signal amplification.

PSA Imaging Kits

PSA imaging kits provide a complete, ready-to-use solution for power Styramide™ signal amplification. Each kit includes an HRP-conjugated secondary antibody, iFluor® Styramide™ reagent, and optimized reaction buffer. These kits are designed for researchers who want a streamlined workflow with validated components.

Key Features of PSA Imaging Kits

Complete kit format with all required reagents
HRP-conjugated secondary antibodies included (anti-mouse, anti-rabbit, or anti-human)
Optimized protocols for consistent results
Multiple fluorophore options covering UV to far-red spectrum

Fig. 3

Sequential immunostaining of formaldehyde-fixed, paraffin-embedded human lung adenocarcinoma using iFluor® PSA imaging kits. EpCam was labeled with rabbit anti-EpCam antibody and iFluor® 488 PSA imaging kit with goat anti-rabbit IgG (Catalog Number 45205), followed by washing. Pan-keratin was labeled with mouse anti-pan keratin antibodies and iFluor® 555 PSA imaging kit with goat anti-mouse IgG (Catalog Number 45270). Nuclei were labeled with DAPI (Catalog Number 17507). Images were acquired on a confocal microscope.

mFluor™ Styramide™ Conjugates

mFluor™ Styramide™ conjugates feature fluorophores with large Stokes shift that are optimized around common laser lines. This property is particularly useful for multiplex imaging applications where spectral overlap is a concern, allowing more markers to be excited by a shared laser but read in different emission channels.

Key Features of mFluor™ Styramide™ Conjugates

Large Stokes shifts (100-300 nm) reduce spectral crosstalk
Common excitation for simplified multiplexing
Compatible with standard filter sets
Ideal for violet and blue laser excitation systems

Hapten-Based Styramide™ Conjugates (Biotin, DIG, DNP)

Hapten-based Styramide™ conjugates provide flexible detection options through secondary detection with anti-hapten antibodies or streptavidin conjugates. These reagents are particularly useful for multiplex applications where different haptens can be detected sequentially or in combination with fluorescent styramides.

Key Features of Hapten-Based Styramide™ Conjugates

Indirect detection allows signal amplification through secondary antibody or streptavidin-based detection reagents
Multiple hapten options (Biotin, DIG, DNP) for flexible experimental design
Enhanced sensitivity compared to traditional tyramide-hapten conjugates
Compatible with standard streptavidin or anti-hapten detection systems

iFluor® and Classic Tyramides (TSA Technology)

Tyramide signal amplification (TSA) reagents use HRP-catalyzed deposition of labeled tyramide molecules for enhanced immunodetection. AAT Bioquest offers both iFluor®-conjugated tyramides as well as classic tyramide formulations including fluorescein, Cy3, and Cy5 conjugates.

Buffers, Accessories, and Supporting Reagents

These supporting products are essential for optimizing TSA and PSA workflows. The ReadiUse™ tyramide (TSA)/Styramide™ (PSA) stripping buffer enables non-thermal removal of antibodies and deposited fluorophores after TSA or PSA staining, allowing sequential multiplex immunofluorescence without heat-induced tissue damage or epitope loss. The ReadiUse™ optimized reaction buffer enhances signal intensity and reduces background, while the Amplite® signal booster further improves detection sensitivity. The FluoroQuest™ TSA/PSA antifade mounting medium is specifically formulated to preserve fluorescence signals from tyramide and Styramide™ staining.

This document (01.0222.251203r1) was last updated on Tue Mar 31 2026. All trademarks and registered trademarks mentioned herein are the property of their respective owners.