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Sudan Black B treatment reduces autofluorescence and improves resolution of in situ hybridization specific fluorescent signals of brain sections.
Oliveira, V C; Carrara, R C V; Simoes, D L C; Saggioro, F P; Carlotti, C G; Covas, D T; Neder, L.
Affiliation
  • Oliveira VC; Department of Pathology, Ribeirão Preto School of Medicine - University of São Paulo (USP), Ribeirão Preto - SP, Brazil. vivianecassia@usp.br
Histol Histopathol ; 25(8): 1017-24, 2010 08.
Article in En | MEDLINE | ID: mdl-20552552
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.
Subject(s)

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Histocytological Preparation Techniques / Coloring Agents Type of study: Guideline Limits: Humans Language: En Journal: Histol Histopathol Journal subject: HISTOLOGIA / PATOLOGIA Year: 2010 Document type: Article Affiliation country: Brasil Country of publication: España

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Histocytological Preparation Techniques / Coloring Agents Type of study: Guideline Limits: Humans Language: En Journal: Histol Histopathol Journal subject: HISTOLOGIA / PATOLOGIA Year: 2010 Document type: Article Affiliation country: Brasil Country of publication: España