Vector® TrueVIEW® Autofluorescence Quenching Kit

SP-8400
SKU Unit Size Price Qty
SP-8400-15 15 ml
$142.00

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Description

The TrueVIEW Autofluorescence Quenching Kit 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
  • TrueVIEW with DAPI counterstain available here

 

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

Specifications

More Information
Unit Size 15 ml
Applications Immunofluorescence, In situ hybridization
Blocking Action Autofluorescence
Format Concentrate

Kit Components

Kit Contents:

  • TrueVIEW Reagent A, 5 ml
  • TrueVIEW Reagent B, 5 ml
  • TrueVIEW Reagent C, 5 ml
  • VECTASHIELD® Vibrance™ Antifade Mounting Medium, 2 ml

Documents

Product FAQs

Why is the TrueVIEW™ Autofluorescence Quenching reagent applied after completion of the IF assay and not at the start of the procedure?

During product development, applying the TrueVIEW™ Quenching reagent at the end of our standard IF procedure yielded the most optimal reduction in autofluorescence signal. The reagent is retained on the tissue at the time of mounting, allowing for extended quenching action, with little to no effect on the specific fluorescent signal. However, in applications with very brief staining procedures, such as a primary antibody directly conjugated with a fluorophore, application of TrueVIEW™ Quenching reagent may be just as effective at the start of the procedure.

Do you have any published references describing the use of TrueVIEW Autofluorescence Quenching Kit?

There are a number of published references describing the use of TrueVIEW Autofluorescence Quenching Kit in the scientific literature. Please see a partial list of publications in the Technical Information section.

Is TrueVIEW effective against lipofuscin derived autofluorescence?

The working solution of TrueVIEW works in an electrostatic manner to greatly reduce or eliminate fluorescence in tissue sections induced through the use of an aldehyde based fixative. TrueVIEW is also effective at reducing fluorescence from tissue components such as collagen, elastin and red blood cells. TureVIEW does not work to reduce autofluorescence due to lipofuscin.

How long can I store the TrueVIEW working solution?

Once made up, the working solution of TrueVIEW can be stored either on the bench top or in the fridge (2-8C) fro about 48 hours (2 days) withut loss of activity or function. Following this time we would suggest discarding unused working solution and make fresh solution as required.

My tissue sections turned blue when I applied TrueVIEW. Is this supposed to happen?

Yes, the working solution is a blue color that does "stain" the tissue section blue. This indicates an active and appropriate chemical reaction is occurring. The blue stain on the section does not fluoresce and does not interfere with the immunofluorescence application.

What mounting media can I use with TrueVIEW?

Both TrueVIEW products, SP-8400 and SP-8500, are supplied with 2 mL of VECTASHIELD Vibrance antifade mounting media. We have found that the mounting media does play a crucial role in maintaining the high signal to noise ratio when using TrueVIEW. VECTASHIELD Vibrance is included therefore as a critical component of the kit and it is recommended to use the media supplied for optimal results. At this time we do not have sufficient data to confidently recommend the use of other vendors mounting media with TrueVIEW. Substitution of VECTASHIELD Vibrance with another mounting media may result in less than satisfactory results.

For the wash step after applying the TrueVIEW quenching reagent, can I use buffers other than PBS or detergents?

No, we have found that the TrueVIEW reagent lifts off the tissue using TBS or HEPES buffer. Detergents are incompatible.

Citations

Technical 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 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, TrueVIEW reagent binds to hydrophilic compounds and effectively quenches endogenous autofluorescence. 

Publications:

  1. McTiernan, C. et al. 2020. LiQD Cornea: Pro-regeneration collagen mimetics as patches and alternatives to corneal transplantation. Science Advances. Vol. 6, no. 25.(https://advances.sciencemag.org/content/6/25/eaba2187)

  2. Hou, Y. et al. 2020. SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract. Cell. 182, 1–18 (https://doi.org/10.1016/j.cell.2020.05.042)

  3. Takenaga, K. et al. Cancer cell-derived interleukin-33 decoy receptor sST2 enhances orthotopic tumor growth in a murine pancreatic cancer model. PLoS ONE. 15(4): e0232230. (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7185704/)

  4. Fiock K. et al. 2020. Increased Tau Expression Correlates with Neuronal Maturation in the Developing Human Cerebral Cortex. eNeuro. 7(3): ENEURO.0058-20.2020. (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7262004/)

  5. Murata, A. and Hayashi, S. 2020. CD4+ Resident Memory T Cells Mediate Long-Term Local Skin Immune Memory of Contact Hypersensitivity in BALB/c Mice. Front Immunol. 11: 775. (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7248184/)
  6. Wilson, M. et al. 2020. Lgr5-positive endothelial progenitor cells occupy a tumor and injury prone niche in the kidney vasa recta. Stem Cell Research. Volume 46, July 2020, 101849. (https://www.sciencedirect.com/science/article/pii/S1873506120301501#!)

  7. Milstone, Z. et al. 2020. Histone deacetylases 1 and 2 silence cryptic transcription to promote mitochondrial function during cardiogenesis. Science Advances. Vol. 6, no. 15, eaax5150 (https://advances.sciencemag.org/content/6/15/eaax5150)

  8. Duler, L. et al. 2020. Identification of Neutrophil Extracellular Traps in Paraffin-Embedded Feline Arterial Thrombi using Immunofluorescence Microscopy. Journal of Visualized Experiments. (157), e60834, doi:10.3791/60834 (https://www.jove.com/video/60834/identification-neutrophil-extracellular-traps-paraffin-embedded?status=a62840k)

  9. Takahashi, K. et al. 2020. Eosinophils are the main cellular targets for oral gene delivery using Lactic acid bacteria. Vaccine. 38(17):3330-3338 (https://www.ncbi.nlm.nih.gov/pubmed/32197922)

  10. Parsons Aubone, A. et al. 2020. Presence of Clock genes in equine full-term placenta. Journal of Animal Science. skaa094 (https://academic.oup.com/jas/advance-article-abstract/doi/10.1093/jas/skaa094/5811452)

  11. Williams, J. et al. 2020. Mylk3 null C57BL/6N mice develop cardiomyopathy, whereas Nnt null C57BL/6J mice do not. Life Sci Alliance. 3(4): e201900593. (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7103425/)

  12. Plant, T. et al. 2020. Semaphorin 3F signaling actively retains neutrophils at sites of inflammation. The Journal of Clinical Investigation. (https://www.jci.org/articles/view/130834)

  13. de Barros Mucci, D. et al. 2020. Impact of maternal obesity on placental transcriptome and morphology associated with fetal growth restriction in mice. International Journal of Obesity. (https://www.nature.com/articles/s41366-020-0561-3)

  14. Cui, H. et al. 2020. Monocyte-derived alveolar macrophage Apolipoprotein E participates in pulmonary fibrosis resolution. JCI Insight. (https://insight.jci.org/articles/view/134539)

  15. Kimura, Y. et al. 2020. Soluble Uric Acid Promotes Atherosclerosis via AMPK (AMP-Activated Protein Kinase)-Mediated Inflammation. Arteriosclerosis, Thrombosis, and Vascular Biology. 40:570–582. (https://www.ahajournals.org/doi/full/10.1161/ATVBAHA.119.313224)

  16. Blanco, S. et al. 2020. Hyaluronate Nanoparticles as a Delivery System to Carry Neuroglobin to the Brain after Stroke. Pharmaceutics  (https://doi.org/10.3390/pharmaceutics12010040)

  17. Fallet, B. et al. 2020. Chronic Viral Infection Promotes Efficient Germinal Center B Cell Responses. Cell Reports. 30, 1013–1026. (https://www.sciencedirect.com/science/article/pii/S2211124719316730#!)

  18. Kerby, A. et al. 2020. Placental Morphology and Cellular Characteristics in Stillbirths in Women With Diabetes and Unexplained Stillbirths. Archives of Pathology & Laboratory Medicine. (https://www.archivesofpathology.org/doi/10.5858/arpa.2019-0524-OA)

  19. Imamura, T. et al. 2020. Insulin deficiency promotes formation of toxic amyloid-β42 conformer co-aggregating with hyper-phosphorylated tau oligomer in an Alzheimer's disease model. Neurobiology of Disease 137:1047392. (https://www.sciencedirect.com/science/article/pii/S0969996120300140)

  20. Bärnthaler, T. et al. 2020. Inhibiting eicosanoid degradation exertsantifibrotic effects in a pulmonary fibrosismouse model and human tissue. Journal of Allergy and Clinical Immunology. Volume 145, Issue 3,, Pages 818-833.e11 (https://www.sciencedirect.com/science/article/pii/S0091674919316252#!)

  21. Vinton, C.L. et al. 2019. Simian Immunodeficiency Virus Infection of Rhesus Macaques Results in Delayed Zika Virus Clearance. mBio (https://doi.org/10.1128/mBio.02790-19)

  22. Ushioda, W. et al. 2019. Neuropathology in Neonatal Mice After Experimental Coxsackievirus B2 Infection Using a Prototype Strain, Ohio-1. Journal of Neuropathology & Experimental Neurology  (https://academic.oup.com/jnen/advance-article-abstract/doi/10.1093/jnen/nlz124/5679985)

  23. Oswald, D.M., Jones, M.B., Cobb, B.A. 2019.  Modulation of hepatocyte sialylation drives spontaneous fatty liver disease and inflammation. Glycobiology (https://academic.oup.com/glycob/advance-article-abstract/doi/10.1093/glycob/cwz096/5628931
  24. Ibrahim, M. et al. 2019. Once Daily Pregabalin Eye Drops for Management of Glaucoma. ACS Nano 2019, 13, 13728−13744. (https://www.sciencedirect.com/science/article/pii/S0969996120300140)

  25. Bencze, J., et al. 2019. Neuropathological characterization of Lemur tyrosine kinase 2 (LMTK2) in Alzheimer’s disease and neocortical Lewy body disease. Scientific Reports (https://www.nature.com/articles/s41598-019-53638-9)

  26. Davies, S.P. et al. 2019. Hepatocytes Delete Regulatory T Cells by Enclysis, a CD4+ T Cell Engulfment Process. Cell Reports (https://www.sciencedirect.com/science/article/pii/S2211124719312653)

  27. Liebers, J. et al. 2019. 3D image analysis reveals differences of CD30 positive cells and network formation in reactive and malignant human lymphoid tissue (classical Hodgkin Lymphoma). PLoS One (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6812863/)

  28. Lecocq, Q. et al. 2019. Noninvasive Imaging of the Immune Checkpoint LAG-3 Using Nanobodies, from Development to Pre-Clinical Use. Biomolecules (https://www.mdpi.com/2218-273X/9/10/548/htm)

  29. Motoike, S. et al. 2019. Clumps of Mesenchymal Stem Cell/Extracellular Matrix Complexes Generated with Xeno-Free Conditions Facilitate Bone Regeneration via Direct and Indirect Osteogenesis. International Journal of Molecular Sciences (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6720767/)

  30. Wilson, M.R. et al. 2019. ARID1A and PI3-kinase pathway mutations in the endometrium drive epithelial transdifferentiation and collective invasion. Nature Communications (https://www.nature.com/articles/s41467-019-11403-6)

  31. Nagai-Okatani, C. et al. 2019. Wisteria floribunda agglutinin staining for the quantitative assessment of cardiac fibrogenic activity in a mouse model of dilated cardiomyopathy. Laboratory Investigation (https://www.nature.com/articles/s41374-019-0279-9)

  32. Sheller-Miller, S. et al. 2019. Cyclic-recombinase-reporter mouse model to determine exosome communication and function during pregnancy. American Journal of Obstetrics and Gynecology (https://www.sciencedirect.com/science/article/abs/pii/S0002937819307744#!)

  33. Abe, H. et al. 2019. Correlation between platelet thrombus formation on collagen-coated beads and platelet aggregation induced by ADP. Transfusion and Apheresis Science (https://www.trasci.com/article/S1473-0502(19)30090-4/abstract)

  34. Singh, B. et al. 2019. Tau is required for progressive synaptic and memory deficits in a transgenic mouse model of α-synucleinopathy. Acta Neuropathologica (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6778173/)

  35. 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)

  36. Rhodes, S. et al. 2019. Cdkn2a (Arf) loss drives NF1-associated atypical neurofibroma and malignant transformation. Human Molecular Genetics (https://academic.oup.com/hmg/article/28/16/2752/5489753)

  37. Su, X. et al. 2019. Imaging and Tracking Stem Cell Engraftment in Ischemic Hearts by Near-Infrared Fluorescent Protein (iRFP) Labeling. Methods in Molecular Biology (https://link.springer.com/protocol/10.1007%2F7651_2019_226)

  38. 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)

  39. 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)

  40. Nishimura, A. et al. 2018. Hypoxia-induced interaction of filamin with Drp1 causes mitochondrial hyperfission–associated myocardial senescence. Science Signaling (https://stke.sciencemag.org/content/11/556/eaat5185.long)

  41. 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)

  42. Soontornniyomkij, V. et al. 2018. Association of antiretroviral therapy with brain aging changes among HIV-infected adults. AIDS (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6115290/)

  43. 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)

  44. Boucher, J.M. et al. 2018. Rab27a regulates human perivascular adipose progenitor cell differentiation. Cardiovasc Drugs Ther. (https://doi.org/10.1007/s10557-018-6813-y)

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
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