Biotium’s new far-red NucSpot 640 nuclear counterstain

Biotium launches first ever mounting media with far-red nuclear counterstain and autofluorescence quenching reagent

Biotium’s new far-red NucSpot 640 nuclear counterstain

Human cerebral cortex cryosections were stained with CF 405S GFAP Antibody (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).

 

Biotium, a leading provider of reagents for immunofluorescence microscopy, is significantly expanding its line of antifade wet-set and hardset EverBrite Mounting Medium with options that include Biotium’s new far-red NucSpot 640 nuclear counterstain. In addition, Biotium is also releasing EverBrite TrueBlack Hardset Mounting Medium for mounting and quenching of lipofuscin autofluorescence in a single step.

Antifade mounting media, like EverBrite, typically includes DAPI as a convenient blue nuclear counterstain for researchers performing immunofluorescence microscopy. However, cross-talk between fluorescence channels and UV-induced photoconversion of DAPI can cause bleed-through fluorescence in neighbouring FITC or Cy 3 channels. Biotium developed EverBrite Mounting Media with NucSpot counterstains as a novel solution to DAPI cross-talk and UV photoconversion, offering bright and specific nuclear imaging with very limited interference in other channels. EverBrite NucSpot 640 is specific for the Cy 5 channel.

Biotium’s TrueBlack lipofuscin quenchers are unique reagents designed to quench lipofuscin autofluorescence commonly found in human and aged animal tissues. New EverBrite TrueBlack Hardset Mounting Medium is the first reagent to offer both mounting and quenching of lipofuscin autofluorescence in a single step for greater convenience. EverBrite TrueBlack Hardset Mounting Medium will be available with DAPI, NucSpot 640, or without a nuclear counterstain.

“Mounting medium has traditionally only provided antifade properties and a convenient DAPI counterstain, but does little to address other issues with fluorescence,” says Lori Roberts PhD, Director of Bioscience at Biotium. “Our new EverBrite products aim to provide both convenience and novel research solutions for our customers.”

This release adds to Biotium’s growing portfolio of novel reagents for immunofluorescence microscopy, offering innovative solutions for all user workflows. Biotium’s new EverBrite with NucSpot and EverBrite TrueBlack Hardset Mounting Medium will be available in a standard 10 mL size or 2 mL trial size. NucSpot 640 nuclear counterstain is currently available with EverBrite Hardset and EverBrite TrueBlack Hardset Mounting Medium. Mounting media variations with NucSpot 680 for the Cy 5.5 channel will be released soon, according to the company.