Support & Resources


Keep it Bright with EverBrite™

High-performance mounting medium for immunofluorescence

Superior antifade mounting media with advanced protection from photobleaching for long-lasting fluorescent signal

Advanced protection from photobleaching, available with and without DAPI. Available wet-set or hardset.
Same high-performance antifade with far-red nuclear counterstain that avoids DAPI crosstalk.
The only mounting medium that quenches lipofuscin background and preserves fluorescent signal.

Choose your 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.

Original EverBrite™ Mounting Medium

Biotium’s EverBrite™ line of mounting media are a revolutionary antifade mounting media optimally formulated for preserving fluorescence of our CF® dyes and other fluorochromes. EverBrite™ Mounting Medium is available in wet-set and hardset formulations with and without DAPI. The EverBrite™ Hardset Mounting Medium hardens completely to form a permanent seal at room temperature in 24 hours. Drop-n-Stain EverBrite™ Mounting Medium provides the wet-set EverBrite™ formulation in a convenient dropper bottle for easy dispensing.

Original EverBrite™ Mounting Medium Features

  • Antifade mounting medium optimally formulated to prevent photobleaching of fluorophores
  • Refractive index is well-matched to that of coverslip glass and immersion oil
  • Available in wet-set or hardset formulations
  • Wet-set formulation available as convenient Drop-n-Stain EverBrite™
  • Available with DAPI, hardest formulation available with NucSpot® 640

Unlike VECTASHIELD® wetset, EverBrite™ does not quench the fluorescence of cyanine-based dyes. Methanol-fixed HeLa cells in a 96-well coversglass bottom dish were stained with mouse anti-tubulin antibody followed by Cy®5 goat anti-mouse secondary antibody, then placed in the indicated mounting medium and imaged by confocal microscopy using the same laser and gain settings. Other widely-used cyanine-based dyes include Cy®3, Alexa Fluor® 555, Alexa Fluor® 647, CF®555, CF®647, CF®660C, and CF®680.
Photostability of Cy®5 staining in HeLa cells with various mounting media. A major advantage of EverBrite™ mounting medium is that, unlike Vectashield®, it does not decrease the fluorescence of cyanine-based fluorophores. Fluorescence values for Cy®5 in Vectashield® media are normalized to PBS time t0 to illustrate the drop in fluorescence of cyanine dyes caused by Vectashield®.

Back to top

EverBrite™ Hardset Mounting Medium with NucSpot®

Clearer counterstaining in the far-red channel

EverBrite™ Hardset with far-red NucSpot® 640 counterstain offers greater nuclear-specificity than propidium iodide (PI) and can avoid problems with cross-talk and photoconversion from DAPI. NucSpot® 640 is a far-red DNA binding dye for the Cy®5 channel.

EverBrite™ with NucSpot® Features

  • Avoid cross-talk and photoconversion from DAPI
  • Superior nuclear specificity compared to PI
  • Also available with EverBrite TrueBlack® Hardset
HeLa cells were fixed with PFA, then stained with CF®488A ConA (green). After staining, cells were permeabilized with 0.1% Triton X-100, then mounted with EverBrite™ Mounting Medium with NucSpot® 640 (magenta).
Rat colon cryosection fixed with 4% PFA, stained with CF®488A phalloidin (actin filaments, green), and mounted with EverBrite™ Hardset Mounting Medium with NucSpot® 640 (nuclei, magenta).

Back to top

EverBrite TrueBlack® Hardset Mounting Medium

All-in-one mounting medium that quenches lipofuscin background, preserves fluorescence signal, and counterstains

Lipofuscin are autofluorescent granules that are a common source of background that make fluorescence imaging virtually impossible in human and aged animal tissues such as brain and retina. EverBrite TrueBlack® Hardset Mounting Medium is the only mounting medium optimally formulated for quenching lipofuscin fluorescence while offering the same protection against photobleaching as our original EverBrite™ Hardset. The mounting medium is available without nuclear stain, with DAPI, or with NucSpot® 640 nuclear counterstains.

EverBrite TrueBlack® Hardset Mounting Medium 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, NucSpot® 640, or without nuclear counterstain
Human cerebral cortex cryosections were stained with CF®405S GFAP Antibody (ASTRO/1974R) (glial cells, cyan), then mounted with EverBrite™ Hardset with NucSpot® 640, or EverBrite TrueBlack® Hardset with NucSpot® 640. Lipofuscin autofluorescence appears orange in the merged image and is effectively quenched after mounting in EverBrite TrueBlack®. Nuclei are stained with NucSpot® 640 (magenta).

Methanol-fixed human cerebral cortex cryosections were stained with CF®488A Anti-GFAP, clone GA5+ASTRO/789 (glial cells, green), then mounted with EverBrite™ Hardset or EverBrite TrueBlack® Hardset mounting medium with NucSpot® 640 (nuclei, magenta). Lipofuscin autofluorescence was imaged in the Cy®3 channel (orange). Sections were imaged after curing overnight at room temperature. EverBriteTrueBlack® Hardset effectively quenched lipofuscin autofluorescence.

Back to top