The TrueVIEW Autofluorescence Quenching Kit with DAPI provides a novel way to diminish unwanted autofluorescence from non-lipofuscin sources and dramatically improve signal-to-noise ratio. It removes unwanted autofluorescence in tissue sections due to aldehyde fixation, red blood cells, and structural elements such as collagen and elastin. The quenching action of the kit reagents provides a clear, unambiguous, “true view” localization of the target antigen.

Features:

• Effective on problematic tissues such as kidney and spleen
• Easy-to-use, one-step method
• Compatible with a wide selection of fluorophores
• Compatible with standard epifluorescence and confocal laser microscopes
• Mounting medium contains DAPI for counterstaining in one step

One kit is sufficient to treat approximately 100 to 150 tissue sections. 

Tissue autofluorescence often occurs with aldehyde fixation or from inherent native tissue components (collagen, elastin, and red blood cells). The extent and intensity of autofluorescence background frequently makes it difficult or impossible to distinguish specific signals in immunofluorescence applications.

Most methods for reduction of tissue autofluorescence act primarily on lipofuscin granules, and are not broadly effective against the most common sources of autofluorescence targeted by TrueVIEW Quencher.

Current methods for reducing autofluorescence primarily include “home brew” concoctions such as sodium borohydride and other ink-based products. These methods are essentially ineffective against aldehyde induced autofluorescence.  In contrast, Vector TrueVIEW reagent binds to hydrophilic compounds and effectively quenches endogenous autofluorescence. 

Human tonsil (FFPE); Section stained for AE1/AE3 using fluorescein label (green). Treated with TrueVIEW and mounted with VECTASHIELD Vibrance Antifade Mounting Medium with DAPI (nuclei blue).

Easy To Apply Following completion of the IF staining procedure:

Mode of Action Following completion of the IF staining procedure:

Signal to Noise Optimization The primary antibody and detection reagents should be optimized (titered) in conjunction with Vector TrueVIEW Quenching Kit to achieve maximum signal to noise.

Optimization of Exposure Increasing exposure times may be necessary to achieve optimal image acquisition for TrueVIEW Quencher treated slides.

Comparison Between Vector TrueVIEW Reagent and Other Autofluorescence Reducing Agents We compared the effectiveness of Vector TrueVIEW quenching action in parallel with other commercially available autofluorescence reducing products and “home brew” reagents, on serial sections of formalin-fixed, paraffin-embedded human pancreas visualized using a standard fluorescein (green) filter. No specific immunofluorescence staining was conducted. The images below highlight our results. All images were acquired under identical conditions (including microscope objective and exposure times).

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