Lipofuscin can make fluorescence staining of human tissues virtually impossible
TrueBlack™ is a new reagent for quenching lipofuscin autofluorescence in tissue sections for immunofluorescence staining. Lipofuscin consists of highly autofluorescent granules of oxidized proteins and lipids that build up in the lysosomes of aging cells. Lipofuscin granules fluoresce brightly in all channels used for fluorescence microscopy, and accumulate in a variety of cell and tissue types with age. Consequently, imaging of specific immunofluorescence signal in certain adult human tissues or aged animal tissues can be virtually impossible unless methods are employed to quench or mask lipofuscin fluorescence.
TrueBlack™ treatment is simple and rapid
TrueBlack™ treatment can be performed before or after immunostaining. It is rapid, simple, and has minimal effect on signal from fluorescent antibodies or nuclear counterstains, thus preserving the signal-to-noise ratio of the immunostaining.
Unlike Sudan Black, TrueBlack™ quenches autofluorescence with minimal background
Traditionally, Sudan Black B has been used to quench lipofuscin autofluorescence by incubating tissue sections with the dye after immunofluorescence staining. However, while it masks the autofluorescence from lipofuscin, Sudan Black B also introduces uniform non-specific background fluorescence in the red and far-red channels, limiting the use of fluorescent dyes in those wavelengths. Biotium developed TrueBlack™ as a superior alternative to Sudan Black B to quench autofluorescence with far less increase in red/far-red background fluorescence.
TrueBlack™ helps reduce autofluorescence from other sources too
TrueBlack effectively eliminates lipofuscin autofluorescence in tissues like human brain and retina. TrueBlack also can reduce autofluorescence from other sources, such as collagen, elastin, red blood cells, and general background fluorescence. It is not as effective at quenching these sources of autofluorescence as it is for lipofuscin, but it can improve background in human and non-human tissue types.
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TrueBlack™ quenches lipofuscin with less background than Sudan Black B
TrueBlack™ treatment can be performed before or after antibody staining
TrueBlack™ can reduce autofluorescence from sources other than lipofuscin
TrueBlack™ also can improve background in non-human tissue