Vector® TrueVIEW™ Autofluorescence Quenching Kit

Cat. No: SP-8400
Price: $137.00
Unit Size: 15 ml

Overview

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
  • TrueVIEW with DAPI counterstain available here

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

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. 

Publications:

  • Boucher, J.M., Robich, M., Scott, S.S. et al (2018). Rab27a Regulates Human Perivascular Adipose Progenitor Cell Differentiation. Cardiovasc Drugs Ther. (https://doi.org/10.1007/s10557-018-6813-y)

  • Du, H. et al (2018). A Novel Mouse Model of Hemangiopericytoma Due to Loss of Tsc2. Human Mol. Gen. (https://doi.org/10.1093/hmg/ddy289)

  • Su Y, Hou Y, Wang Q, (2018). The Enhanced Replication of an S-intact PEDV During Coinfection with an S1 NTD-del PEDV in Piglets, Veterinary Microbiology (https://doi.org/10.1016/j.vetmic.2018.11.025)

  • Yoon, J.H., Li, M., Basile, J.R., Lin, Y (2018). Computer-assisted Analysis of Immunohistological Parameters in Oral Giant Cell Granulomas, Oral Diseases (https://doi.org/10.1111/odi.13022)

  • Rodgers, H.M. et al. (2019). Dopamine D1 and D3 Receptor Modulators Restore Morphine Analgesia and Prevent Opioid Preference in a Model of Neuropathic Pain. Neuroscience (https://www.sciencedirect.com/science/article/abs/pii/S0306452219301927)

  • Chafe, S.C. et al. (2019). Targeting Hypoxia-Induced Carbonic Anhydrase IX Enhances Immune-Checkpoint Blockade Locally and Systemically. Cancer Immunol Res (http://cancerimmunolres.aacrjournals.org/content/early/2019/05/31/2326-6066.CIR-18-0657)

Spleen with-without TrueVIEW

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

 

Spleen w-wo TrueVIEW Quencher Treatment

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:

TrueVIEW Quencher How To

 

Mode of Action

Following completion of the IF staining procedure:

TrueVIEW Quencher mode of action

 

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. 

TrueVIEW Primary Ab Optimization

 

Optimization of Exposure

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

TrueView Optimization of Exposure

 

 

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). 

TrueVIEW Comparisons
TrueVIEW Time Course

 

 Customer Testimonials 

TrueVIEW Testimonials

Safety Information

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