Publication: Sudan Black B treatment reduces autofluorescence
and improves resolution of in situ hybridization
specific fluorescent signals of brain sections
Authors
Oliveira, V.C. ; Carrara, R.C.V. ; Simoes, D.L.C. ; Saggioro, F.P. ; Carlotti, C.G. ; Covas, D.T., Jr. ; Neder, L.
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Publisher
Murcia: F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
Interference by autofluorescence is one of the
major concerns of immunofluorescence analysis of in
situ hybridization-based diagnostic assays. We present a
useful technique that reduces autofluorescent
background without affecting the tissue integrity or
direct immunofluorescence signals in brain sections.
Using six different protocols, such as ammonia/ethanol,
Sudan Black B (SBB) in 70% ethanol, photobleaching
with UV light and different combinations of them in
both formalin-fixed paraffin-embedded and frozen
human brain tissue sections, we have found that tissue
treatment of SBB in a concentration of 0.1% in 70%
ethanol is the best approach to reduce/eliminate tissue
autofluorescence and background, while preserving the
specific fluorescence hybridization signals. This strategy
is a feasible, non-time consuming method that provides a
reasonable compromise between total reduction of the
tissue autofluorescence and maintenance of specific
fluorescent labels.
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