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Characterizing and diminishing autofluorescence in formalin‐fixed paraffin‐embedded human respiratory tissue (LB482)
Author(s) -
Davis Anne,
Richter Anke,
Becker Steven,
Moyer Jenna,
Sandouk Aline,
Skinner Jeff,
Taubenberger Jeffery
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.lb482
Subject(s) - autofluorescence , pathology , antigen retrieval , chemistry , staining , medicine , fluorescence , physics , quantum mechanics
Tissue autofluorescence frequently hampers visualization of immunofluorescent markers in formalin‐fixed paraffin‐embedded respiratory tissues. We assessed nine treatments reported to have efficacy in reducing autofluorescence in other tissue types. The three most efficacious were eriochrome black T, Sudan black B and sodium borohydride. We then further characterized these changes in autofluorescence using white light laser confocal multi‐lambda analysis. Additionally, we assessed the impact of steam antigen retrieval and serum application on human tracheal tissue autofluorescence. Functionally fitting this multi‐lambda data to 2‐dimensional Gaussian surfaces revealed that steam antigen retrieval and serum application contribute minimally to autofluorescence and that the three treatments are disparately efficacious. Together these studies provide a set of guidelines for diminishing autofluorescence in formalin‐fixed paraffin‐embedded human respiratory tissue. Additionally, the characterization techniques are transferable to similar questions in other tissue types as demonstrated on frozen human liver tissue and paraffin embedded mouse lung fixed in different fixatives.