Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue

Michael K. Axthelm; Steven Krakowka

doi:10.1016/0166-0934(86)90016-9

Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue

Michael K. Axthelm, Steven Krakowka

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

The effects of enzymatic digestion, sodium borohydride reduction, acids used in décalcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.

Original language	English (US)
Pages (from-to)	215-229
Number of pages	15
Journal	Journal of Virological Methods
Volume	13
Issue number	3
DOIs	https://doi.org/10.1016/0166-0934(86)90016-9
State	Published - Jun 1986
Externally published	Yes

Keywords

aldehyde fixation
canine distemper virus
immunocytochemistry
immunofluorescence technique
immunoperoxidase
technique

ASJC Scopus subject areas

Virology

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@article{776faff7f6af452fa6629c1a3491df57,

title = "Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue",

abstract = "The effects of enzymatic digestion, sodium borohydride reduction, acids used in d{\'e}calcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.",

keywords = "aldehyde fixation, canine distemper virus, immunocytochemistry, immunofluorescence technique, immunoperoxidase, technique",

author = "Axthelm, {Michael K.} and Steven Krakowka",

note = "Funding Information: The authors thank Nancy Austin, Judy Dubena and Alice Kelley for technical assistance and Virginia Stump for preparation of the manuscript. This work was supported by The State of Ohio Canine Research Fund, The National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health Grant SROl-NS14821-15 and U.S. Public Health Service Grant 2 T32 NS07091-06Al 0021. Copyright: Copyright 2014 Elsevier B.V., All rights reserved.",

year = "1986",

month = jun,

doi = "10.1016/0166-0934(86)90016-9",

language = "English (US)",

volume = "13",

pages = "215--229",

journal = "Journal of Virological Methods",

issn = "0166-0934",

publisher = "Elsevier",

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TY - JOUR

T1 - Immunocytochemical methods for demonstrating canine distemper virus antigen in aldehyde-fixed paraffin-embedded tissue

AU - Axthelm, Michael K.

AU - Krakowka, Steven

N1 - Funding Information: The authors thank Nancy Austin, Judy Dubena and Alice Kelley for technical assistance and Virginia Stump for preparation of the manuscript. This work was supported by The State of Ohio Canine Research Fund, The National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health Grant SROl-NS14821-15 and U.S. Public Health Service Grant 2 T32 NS07091-06Al 0021. Copyright: Copyright 2014 Elsevier B.V., All rights reserved.

PY - 1986/6

Y1 - 1986/6

N2 - The effects of enzymatic digestion, sodium borohydride reduction, acids used in décalcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.

AB - The effects of enzymatic digestion, sodium borohydride reduction, acids used in décalcification procedures and techniques for inactivation of endogenous peroxidase were sequentially evaluated for their effect on the immunoreactivity of canine distemper virus in aldehyde-fixed paraffin-embedded tissue. Enzyme digestion improved immunoreactivity while sodium borohydride reduced background staining. Paraformaldehyde-glutaraldehyde-fixed tissues required thioglycolic acid treatment prior to enzyme digestion and sodium borohydride reduction to obtain results comparable to results obtained in formalin-fixed tissues. Detailed protocols for indirect immunofluorescence and the avidin-biotin-peroxidase complex procedure are provided.

KW - aldehyde fixation

KW - canine distemper virus

KW - immunocytochemistry

KW - immunofluorescence technique

KW - immunoperoxidase

KW - technique

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JF - Journal of Virological Methods

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