Aluora™ Spatial Amplification Individual Dyes

Aluora™ Spatial Amplification individual dyes enable a highly sensitive method for multiplex immunohistochemical (mIHC) analysis for use and in-situ hybridization (ISH) analysis for use with the Aluora HRP reagents. These dyes enable bright fluorescent covalent marker labeling with precision and sensitivity 10–200X greater than standard IHC and 2–10X that of other tyramide and CARD techniques.

Revolutionary signal amplification technology

The Aluora spatial amplification kits represent an advance in signal amplification for spatial proteomics and transcriptomics. They employ proprietary Aluora reagents that react with horseradish peroxidase (HRP) to deposit photostable Aluora dyes onto target proteins, significantly improving the signal-to-background and helping reduce the impact of tissue autofluorescence in commonly imaged wavelengths. This chemistry enables robust multiplexed marker detection even in challenging tissue samples. Aluora kits requires 10–5000X less primary antibody than standard and multiplex IHC with antibodies alone and achieve greater or equal signal intensity.

Aluora spatial amplification reagents are optimized for use with Prolong Glass Antifade mountants and 1 mg/mL Thermo Scientific DAPI nucleic acid stains for multiplex analysis.

Aluora reagents and dyes can be purchased individually for use in IHC and ISH workflows or in pre-configured kits to detect mouse, rabbit, or biotin-labelled antibodies. The Aluora multiplex kits come as dye-only kits configured with 3 to 8 dyes or as the Aluora rainbow kit is configured with 8 Aluora dyes and 4X GAM and 4X GAR secondary detection core reagents for 100 slides in 8-plex.

Features of Aluora spatial amplification reagents include:

• Signal amplification for enhanced detection sensitivity
• Comprehensive spatial analysis and imaging capabilities for research applications
• Fluorescent detection system for multiplexing and quantitative analysis
• Flexible workflow options that accommodate diverse experimental needs
• Polymerase HRP (PolyHRP) conjugated secondary antibodies that boost signal for low abundance targets
• Highly cross-adsorbed secondary antibodies that help ensure target-specific detection and background reduction
• Enhanced background reduction through proprietary blocking reagents for crisp, clear imaging

Optimized for automated slide staining

A major benefit of Aluora spatial amplification kits is their compatibility with automated slide stainers right out of the box. This enables complete labeling of a tissue slide with an 8-plex Aluora panel and DAPI with walk-away automation sample processing.

Reducing background for cleaner images

Aluora spatial amplification kits include blockers to help reduce endogenous peroxidase activity and autofluorescence. By addressing autofluorescence and background noise, less primary antibody is needed to generate a bright specific signal.

Iterative labeling enables unlimited multiplexing

The covalently bound Aluora dyes are resistant to antibody stripping, remaining brightly fluorescent through subsequent labeling cycles. This enables iterative staining of numerous targets using primary antibodies from the same host species—a major constraint in traditional immunohistochemistry. Any number of directly conjugated primary antibodies can also be introduced after Aluora labeling is complete.

Custom optimization for diverse applications

Although general protocols are provided, each sample requires optimization for optimal results. Tissue preparation and processing methods vary widely and can profoundly impact staining. Careful optimization of the complete workflow is critical to achieve the highest quality multiplexed data from precious samples. Aluora spatial amplification provides a versatile tool to address diverse spatial proteomics applications.