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EverBrite TrueBlack® Hardset Mounting Medium

Unique hardset antifade that quenches lipofuscin autofluorescence. Choose with mounting medium with DAPI, far-red NucSpot® 640, or without nuclear stain.

Product Attributes

For live or fixed cells

For fixed cells/tissue sections

Probe cellular localization

Nucleus

Excitation/Emission

DAPI: 358/461 nm (with DNA), NucSpot® 640: 649/668 nm (with DNA)

Storage Conditions

Store at 2 to 8 °C, Protect from light

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2 mL
10 mL
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Product Description

This unique hardset antifade mounting medium quenches lipofuscin autofluorescence, for one-step mounting, quenching, and optional nuclear counterstaining of fluorescently-stained tissue sections.

Features

  • The only mounting medium with autofluorescence quenching
  • Quenches as it hardens, with low background fluorescence
  • Optimally formulated for protecting CF® Dyes and other dyes from photobleaching
  • Refractive index well-matched to coverglass (1.46 after curing)
  • Choice of DAPI, far-red NucSpot® 640, or without nuclear counterstain

Spectral Properties of Nuclear Stains

  • DAPI: 358/461 (with DNA)
  • NucSpot® 640: 649/668 nm (with DNA) (for Cy®5 channel)

TrueBlack® Lipofuscin Quenchers are designed to quench lipofuscin autofluorescence, a common source of background in human and aged animal tissues such as brain and retina. They also may reduce background from other sources like red blood cells or extracellular matrix. EverBrite TrueBlack® Hardset Mounting Medium performs lipofuscin quenching and mounting in a single step, for greater convenience. The mounting medium substantially reduces lipofuscin fluorescence after mounting, and quenching continues to improve as the medium cures and hardens overnight. Unlike VECTASHIELD® Mounting Medium, EverBrite TrueBlack® is compatible with cyanine-based fluorophores and is optimally formulated for use with Biotium’s CF® Dyes.

Note: EverBrite TrueBlack® Hardset may reduce specific fluorescence; antibody or probe concentration may require optimization for use with the mounting medium.

EverBrite TrueBlack® is available with or without DAPI, or with Biotium’s novel NucSpot® nuclear-specific counterstains. NucSpot® 640 is a far-red dye for the Cy®5 channel. Using a far-red nuclear stain can avoid problems of fluorescence cross-talk and UV photoconversion with traditional dyes like DAPI. See our other NucSpot® Nuclear Stains for bright and specific nuclear counterstaining, available from green to near-IR.

Note: NucSpot® dyes also show dim blue fluorescence with the DAPI filter set, and should be tested for suitability before using with blue probes, especially for epifluorescence microscopy.

Choose the right quencher for your application

ProductCatalog no.ProsCons
TrueBlack® Lipofuscin Autofluorescence Quencher23007• Complete quenching of lipofuscin autofluorescence

• Ultra-low background in blue and green channels

• Quenching takes only 30 seconds
• Introduces some red/far-red background

• Quenching performed in 70% EtOH

• Some quenching of fluorescent dyes
TrueBlack® Plus Lipofuscin Autofluorescence Quencher23014• Quenching performed in PBS or other buffer

• Greatly reduces lipofuscin autofluorescence

• Has lower red/far-red background than the original TrueBlack®

• Titration recommended for optimal quenching

• May not be as effective as the original TrueBlack® for high-lipofuscin samples

• Some quenching of fluorescent dyes
EverBrite TrueBlack® Hardset Mounting Medium23017• The only mounting medium for one-step mounting and quenching

• Greatly reduces lipofuscin autofluorescence

• Has lower red/far-red background than the original TrueBlack®

• Choice of nuclear counterstains
• May not be as effective as the original TrueBlack® for high-lipofuscin samples

• Some quenching of fluorescent dyes

• NucSpot® dyes should be tested for suitability with blue dyes
EverBrite TrueBlack® Hardset Mounting Medium with DAPI23018
EverBrite TrueBlack® Hardset Mounting Medium with NucSpot® 64023019

See our full selection of antifade mounting media:

ProductNuclear CounterstainCat. No.Features
EverBrite™ Mounting MediumNone23001• Wet-set mounting medium
• Requires coverslip sealing
• Refractive index 1.46
EverBrite™ with DAPIDAPI23002
Drop-n-Stain EverBrite™ Mounting MediumNone23008• Wet-set mounting medium
• Convenient dropper bottle
• Ideal for wells & chambers
• Refractive index 1.42
Drop-n-Stain EverBrite™ with DAPIDAPI23009
EverBrite™ Hardset Mounting MediumNone23003• Hard-set mounting medium
• Forms hard seal after 24 h
• No coverslip sealing needed
• Refractive index 1.46 after curing
EverBrite™ Hardset with DAPIDAPI23004
EverBrite™ Hardset with NucSpot® 640NucSpot® 64023016
EverBrite TrueBlack® Hardset Mounting MediumNone23017• Unique antifade with lipofuscin quenching
• Quenches as it hardens, with low background
EverBrite TrueBlack® Hardset with DAPIDAPI23018
EverBrite TrueBlack® Hardset with NucSpot® 640NucSpot® 64023019
CoverGrip™ Coverslip SealantN/A23005• For sealing edges of wet-set coverslips
* Note: NucSpot® 640 also have dim fluorescence with the DAPI filter set, and should be tested for suitability with blue probes, especially for epifluorescence imaging.
Cy Dye is a registered trademark of Cytiva; VECTASHIELD is a registered trademark of Vector Laboratories.
 

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