Selection of a fixative for identifying T cell subsets, B cells, and macrophages in paraffin-embedded mouse spleen

Howard Eliot Gendelman, Thomas R. Moench, Opendra Narayan, Diane E. Griffin

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Fixation techniques were investigated for their ability to preserve morphology, esterase activity and cell surface antigens in paraffin-embedded mouse lymphoid tissue. The avidin-biotin-peroxidase system was used to stain antigens Thy-1.2, Lyt-1, Lyt-2, RA32C2 and F4/80. Conventional fixatives were compared with fixatives containing periodate and lysine plus paraformaldehyde and/or glutaraldehyde. Conventional fixatives preserved esterase activity but not many cell surface antigens. Periodate-lysine fixatives allowed better preservation of membrane antigens, but esterase activity was often lost at antigen-preserving concentrations of paraformaldehyde or glutaraldehyde. However, a periodate-lysine fixative containing both paraformaldehyde and glutaraldehyde preserved esterase and showed good to excellent staining of Lyt-1, Thy-1.2, RA32C2, and F4/80. Lyt-2 could not be stained with any fixative, but was well preserved in frozen material post-fixed with periodate-lysine based fixatives. We conclude that with proper fixation immune cell surface markers can be identified in paraffin-embedded tissue.

Original languageEnglish (US)
Pages (from-to)137-145
Number of pages9
JournalJournal of Immunological Methods
Volume65
Issue number1-2
DOIs
StatePublished - Dec 16 1983

Fingerprint

Fixatives
T-Lymphocyte Subsets
Paraffin
B-Lymphocytes
Spleen
Macrophages
Esterases
Glutaral
Lysine
Surface Antigens
Thy-1 Antigens
Antigens
Avidin
Lymphoid Tissue
Biotin
Peroxidase
Coloring Agents
Staining and Labeling
Membranes

Keywords

  • cell surface antigen
  • immunoperoxidase staining
  • monoclonal antibody
  • paraffin embedding
  • periodate-lysine-paraformaldehyde-glutaraldehyde

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Selection of a fixative for identifying T cell subsets, B cells, and macrophages in paraffin-embedded mouse spleen. / Gendelman, Howard Eliot; Moench, Thomas R.; Narayan, Opendra; Griffin, Diane E.

In: Journal of Immunological Methods, Vol. 65, No. 1-2, 16.12.1983, p. 137-145.

Research output: Contribution to journalArticle

@article{5bb992b46dbc4d5bac09e90c08e2e26a,
title = "Selection of a fixative for identifying T cell subsets, B cells, and macrophages in paraffin-embedded mouse spleen",
abstract = "Fixation techniques were investigated for their ability to preserve morphology, esterase activity and cell surface antigens in paraffin-embedded mouse lymphoid tissue. The avidin-biotin-peroxidase system was used to stain antigens Thy-1.2, Lyt-1, Lyt-2, RA32C2 and F4/80. Conventional fixatives were compared with fixatives containing periodate and lysine plus paraformaldehyde and/or glutaraldehyde. Conventional fixatives preserved esterase activity but not many cell surface antigens. Periodate-lysine fixatives allowed better preservation of membrane antigens, but esterase activity was often lost at antigen-preserving concentrations of paraformaldehyde or glutaraldehyde. However, a periodate-lysine fixative containing both paraformaldehyde and glutaraldehyde preserved esterase and showed good to excellent staining of Lyt-1, Thy-1.2, RA32C2, and F4/80. Lyt-2 could not be stained with any fixative, but was well preserved in frozen material post-fixed with periodate-lysine based fixatives. We conclude that with proper fixation immune cell surface markers can be identified in paraffin-embedded tissue.",
keywords = "cell surface antigen, immunoperoxidase staining, monoclonal antibody, paraffin embedding, periodate-lysine-paraformaldehyde-glutaraldehyde",
author = "Gendelman, {Howard Eliot} and Moench, {Thomas R.} and Opendra Narayan and Griffin, {Diane E.}",
year = "1983",
month = "12",
day = "16",
doi = "10.1016/0022-1759(83)90310-1",
language = "English (US)",
volume = "65",
pages = "137--145",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Selection of a fixative for identifying T cell subsets, B cells, and macrophages in paraffin-embedded mouse spleen

AU - Gendelman, Howard Eliot

AU - Moench, Thomas R.

AU - Narayan, Opendra

AU - Griffin, Diane E.

PY - 1983/12/16

Y1 - 1983/12/16

N2 - Fixation techniques were investigated for their ability to preserve morphology, esterase activity and cell surface antigens in paraffin-embedded mouse lymphoid tissue. The avidin-biotin-peroxidase system was used to stain antigens Thy-1.2, Lyt-1, Lyt-2, RA32C2 and F4/80. Conventional fixatives were compared with fixatives containing periodate and lysine plus paraformaldehyde and/or glutaraldehyde. Conventional fixatives preserved esterase activity but not many cell surface antigens. Periodate-lysine fixatives allowed better preservation of membrane antigens, but esterase activity was often lost at antigen-preserving concentrations of paraformaldehyde or glutaraldehyde. However, a periodate-lysine fixative containing both paraformaldehyde and glutaraldehyde preserved esterase and showed good to excellent staining of Lyt-1, Thy-1.2, RA32C2, and F4/80. Lyt-2 could not be stained with any fixative, but was well preserved in frozen material post-fixed with periodate-lysine based fixatives. We conclude that with proper fixation immune cell surface markers can be identified in paraffin-embedded tissue.

AB - Fixation techniques were investigated for their ability to preserve morphology, esterase activity and cell surface antigens in paraffin-embedded mouse lymphoid tissue. The avidin-biotin-peroxidase system was used to stain antigens Thy-1.2, Lyt-1, Lyt-2, RA32C2 and F4/80. Conventional fixatives were compared with fixatives containing periodate and lysine plus paraformaldehyde and/or glutaraldehyde. Conventional fixatives preserved esterase activity but not many cell surface antigens. Periodate-lysine fixatives allowed better preservation of membrane antigens, but esterase activity was often lost at antigen-preserving concentrations of paraformaldehyde or glutaraldehyde. However, a periodate-lysine fixative containing both paraformaldehyde and glutaraldehyde preserved esterase and showed good to excellent staining of Lyt-1, Thy-1.2, RA32C2, and F4/80. Lyt-2 could not be stained with any fixative, but was well preserved in frozen material post-fixed with periodate-lysine based fixatives. We conclude that with proper fixation immune cell surface markers can be identified in paraffin-embedded tissue.

KW - cell surface antigen

KW - immunoperoxidase staining

KW - monoclonal antibody

KW - paraffin embedding

KW - periodate-lysine-paraformaldehyde-glutaraldehyde

UR - http://www.scopus.com/inward/record.url?scp=0021042295&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021042295&partnerID=8YFLogxK

U2 - 10.1016/0022-1759(83)90310-1

DO - 10.1016/0022-1759(83)90310-1

M3 - Article

VL - 65

SP - 137

EP - 145

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

IS - 1-2

ER -