Removes unwanted autofluorescence in tissue sections due to aldehyde fixation, red blood cells, and structural elements such as collagen and elastin
Improves overall signal-to-noise in most immunofluorescence assays
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
Kit Contents:
Vector TrueVIEW Reagent A, 5 ml
Vector TrueVIEW Reagent B, 5 ml
Vector TrueVIEW Reagent C, 5 ml
VECTASHIELD® Vibrance™ Antifade Mounting Medium, 2 ml
One kit is sufficient to treat approximately 100 to 150 tissue sections.
Cat. No.
SP-8400-15
Unit Size
15 ml
Country of Manufacture
United States
Applications
Immunofluorescence, In situ hybridization
Blocking Action
Autofluorescence
Format
Concentrate
More Information
Features:
Removes unwanted autofluorescence in tissue sections due to aldehyde fixation, red blood cells, and structural elements such as collagen and elastin
Improves overall signal-to-noise in most immunofluorescence assays
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
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 Vector TrueVIEW Quencher.
The Vector TrueVIEW Autofluorescence Quenching Kit provides a novel way to diminish unwanted autofluorescence from non-lipofuscin sources and dramatically improve signal-to-noise ratio. The quenching action of the kit reagents provides a clear, unambiguous, “true view” localization of the target antigen.
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.
Kit Contents:
Vector TrueVIEW Reagent A, 5 ml
Vector TrueVIEW Reagent B, 5 ml
Vector TrueVIEW Reagent C, 5 ml
VECTASHIELD Vibrance Antifade Mounting Medium, 2 ml
One kit is sufficient to treat approximately 100 to 150 tissue sections.
Spleen (FFPE), antigen retrieved with Antigen Unmasking Solution, stained using Mouse Anti-CD20 (red) and Rabbit Anti-Ki67 (green), followed by VectaFluor™ Duet Kit, and mounted in VECTASHIELD Vibrance Mounting Medium with DAPI. Left: No TrueVIEW Quencher treatment - note interfereing autofluorescence. Right: Treated with TrueVIEW Autofluorescence Quencher.
No TrueVIEW Quencher => 20x objective, red channel exposed 200ms, green channel 200ms
With TrueVIEW Quencher => 20x objective, red channel exposed 200ms, green channel exposed 800ms
Pancreas (FFPE), antigen retrieved with Antigen Unmasking Solution, stained with Guinea Pig x Insulin (green) and Mouse x END (red), followed by FL-Goat x Guinea Pig IgG + Dylight®594-Horse x Mouse IgG. Mounted in VECTASHIELD Vibrance Mounting Medium with DAPI.
No TrueVIEW Quencher => 20x objective, red channel exposed 600ms, green channel 600ms
With TrueVIEW Quencher => 20x objective, red channel exposed 600ms, green channel exposed 600ms
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).
