This unique reagent quenches lipofuscin autofluorescence in tissue sections for immunofluorescence staining and can also help to reduce autofluorescence from other sources like extracellular matrix.
TrueBlack Lipofuscin Autofluorescence Quencher, 20X in DMF, is a novel reagent that quenches lipofuscin autofluorescence in tissue sections for immunofluorescence staining.
- Eliminates autofluorescence of lipofuscin
- Reduces autofluorescence from sources other than lipofuscin
- Does not cause a high record, unlike Sudan Black B
- Can be used before or after immunofluorescence staining
- Clears the way for fluorescence imaging of aged human and animal tissues
TrueBlack effectively removes lipofuscin autofluorescence in tissues such as the human and mouse brain and retina (see Figures 1-3 and TrueBlack references). Lipofuscin consists of highly autofluorescent granules of oxidized proteins and lipids that accumulate in the lysosomes of aging cells.
Lipofuscin granules exhibit intense fluorescence in all channels used for fluorescence microscopy and accumulate in a variety of cell and tissue types with age.
Consequently, imaging of the specific immunofluorescence signal in certain adult human tissues or aged animal tissues can be virtually impossible unless methods are employed to quench or mask the fluorescence of lipofuscin.
TrueBlack turns off lipofuscin with less track record than Sudan Black B
Traditionally, Sudan Black B has been used to quench lipofuscin autofluorescence by incubating tissue sections with the dye after immunofluorescence staining. However, although it masks the autofluorescence of lipofuscin, Sudan Black B also introduces a nonspecific uniform background fluorescence in the red and far-red channels (Fig. 1), limiting the use of fluorescent dyes at these wavelengths.
Now Biotium has developed TrueBlack as a superior alternative to Sudan Black B. TrueBlack quenches lipofuscin fluorescence with a much lower increase in red/far red background fluorescence (Fig. 1). The TrueBlack treatment can be performed before or after immunostaining. It is fast, simple, and has minimal effect on the signal from fluorescent antibodies or nuclear counterstains, thus preserving the signal-to-noise ratio of immunostaining (Figs. 2-3).
TrueBlack can reduce autofluorescence from sources other than lipofuscin
TrueBlack can also reduce autofluorescence from other sources, such as collagen, elastin, red blood cells, and general background fluorescence. It is not as effective in turning off these sources of autofluorescence as it is for lipofuscin, but it can enhance background in human and non-human tissue types (Figs. 4-5). TrueBlack has also been used to quench background fluorescence in polycarbonate filters used as cell supports for image cytometry.
TrueBlack Lipofuscin Autofluorescence Quencher belongs to our TrueBlack line of background reducing agents for fluorescence applications, which includes the TrueBlack WB blocking buffer kit for Western blotting and the TrueBlack® IF background suppressor system (permeabilizer) to block the non-specific immunofluorescence staining. See also our other accessory products for immunofluorescence staining and other applications.