Characterizing and Diminishing Autofluorescence in Formalin-fixed Paraffin-embedded Human Respiratory Tissue
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Authors
Davis, A. Sally
Richter, Anke
Becker, Steven
Moyer, Jenna E.
Sandouk, Aline
Skinner, Jeff
Taubenberger, Jeffery K.
Subjects
autofluorescence
formalin-fixation
paraffin-embedded tissue
immunohistochemistry
immunofluorescence
human
respiratory
confocal microscopy
formalin-fixation
paraffin-embedded tissue
immunohistochemistry
immunofluorescence
human
respiratory
confocal microscopy
Advisors
Date of Issue
2014
Date
Publisher
Language
Abstract
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, as measured using
white light laser confocal Ʌ² (multi-lambda) analysis. We also assessed the impact of steam antigen retrieval and serum
application on human tracheal tissue autofluorescence. Functionally fitting this Ʌ² 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, these characterization techniques are transferable
to similar questions in other tissue types, as demonstrated on frozen human liver tissue and paraffin-embedded mouse lung
tissue fixed in different fixatives.
Type
Article
Description
The article of record as published may be found at http://dx.doi.org/10.1369/0022155414531549
Series/Report No
Department
Organization
Naval Postgraduate School (U.S.)
Identifiers
NPS Report Number
Sponsors
Funder
NIH
NIAID
NIAID
Format
Citation
Davis, A. Sally, et al. "Characterizing and diminishing autofluorescence in formalin-fixed paraffin-embedded human respiratory tissue." Journal of Histochemistry & Cytochemistry 62.6 (2014): 405-423.
Distribution Statement
Rights
This publication is a work of the U.S. Government as defined in Title 17, United States Code, Section 101. Copyright protection is not available for this work in the United States.