Introduction of Zinc-salt Fixation for Effective Detection of Immune Cell-related Markers by Immunohistochemistry

Hidetoshi Mori, Pan Soonsawad, Louis Schuetter, Qian Chen, Neil Hubbard, Robert Cardiff, Alexander D Borowsky

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Tissue localization of immune cells is critical to the study of disease processes in mouse models of human diseases. However, immunohistochemistry (IHC) for immune cell phenotyping in mouse tissue sections presents specific technical challenges. For example, CD4 and CD8 have been difficult to detect using IHC on formalin-fixed and paraffin-embedded mouse tissue, prompting alternative methods. We investigated the use of formalin-free zinc-salt fixation (ZN) and optimized IHC protocols for detecting a panel of immune cell-related markers (CD3, CD4, CD8, Foxp3, B220, F4/80, CD68, and major histocompatibility complex [MHC] class-I, MHC class-II, and Gr-1). The IHC results for these markers were compared on mouse spleen tissue treated with neutral buffered formalin (NBF) or ZN with or ZN without antigen retrieval (AR). Whereas CD4 and CD8 were not detected in NBF-treated tissue, all markers were detected in ZN-treated tissue without AR. Thus, the use of ZN treatment for IHC staining can be a good tool for studying immunoreactive lesions in tissues.

Original languageEnglish (US)
Pages (from-to)883-889
Number of pages7
JournalToxicologic Pathology
Volume43
Issue number6
DOIs
StatePublished - Aug 5 2015

Fingerprint

Zinc
Salts
Immunohistochemistry
Tissue
Formaldehyde
Major Histocompatibility Complex
Antigens
Paraffin
Spleen
Staining and Labeling

Keywords

  • antigen retrieval
  • formalin fixation
  • immune cells
  • immunohistochemistry
  • zinc-salt fixation

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Toxicology
  • Cell Biology
  • Molecular Biology

Cite this

Introduction of Zinc-salt Fixation for Effective Detection of Immune Cell-related Markers by Immunohistochemistry. / Mori, Hidetoshi; Soonsawad, Pan; Schuetter, Louis; Chen, Qian; Hubbard, Neil; Cardiff, Robert; Borowsky, Alexander D.

In: Toxicologic Pathology, Vol. 43, No. 6, 05.08.2015, p. 883-889.

Research output: Contribution to journalArticle

@article{488477baa28d46a78e79bdef3283d585,
title = "Introduction of Zinc-salt Fixation for Effective Detection of Immune Cell-related Markers by Immunohistochemistry",
abstract = "Tissue localization of immune cells is critical to the study of disease processes in mouse models of human diseases. However, immunohistochemistry (IHC) for immune cell phenotyping in mouse tissue sections presents specific technical challenges. For example, CD4 and CD8 have been difficult to detect using IHC on formalin-fixed and paraffin-embedded mouse tissue, prompting alternative methods. We investigated the use of formalin-free zinc-salt fixation (ZN) and optimized IHC protocols for detecting a panel of immune cell-related markers (CD3, CD4, CD8, Foxp3, B220, F4/80, CD68, and major histocompatibility complex [MHC] class-I, MHC class-II, and Gr-1). The IHC results for these markers were compared on mouse spleen tissue treated with neutral buffered formalin (NBF) or ZN with or ZN without antigen retrieval (AR). Whereas CD4 and CD8 were not detected in NBF-treated tissue, all markers were detected in ZN-treated tissue without AR. Thus, the use of ZN treatment for IHC staining can be a good tool for studying immunoreactive lesions in tissues.",
keywords = "antigen retrieval, formalin fixation, immune cells, immunohistochemistry, zinc-salt fixation",
author = "Hidetoshi Mori and Pan Soonsawad and Louis Schuetter and Qian Chen and Neil Hubbard and Robert Cardiff and Borowsky, {Alexander D}",
year = "2015",
month = "8",
day = "5",
doi = "10.1177/0192623315587593",
language = "English (US)",
volume = "43",
pages = "883--889",
journal = "Toxicologic Pathology",
issn = "0192-6233",
publisher = "SAGE Publications Inc.",
number = "6",

}

TY - JOUR

T1 - Introduction of Zinc-salt Fixation for Effective Detection of Immune Cell-related Markers by Immunohistochemistry

AU - Mori, Hidetoshi

AU - Soonsawad, Pan

AU - Schuetter, Louis

AU - Chen, Qian

AU - Hubbard, Neil

AU - Cardiff, Robert

AU - Borowsky, Alexander D

PY - 2015/8/5

Y1 - 2015/8/5

N2 - Tissue localization of immune cells is critical to the study of disease processes in mouse models of human diseases. However, immunohistochemistry (IHC) for immune cell phenotyping in mouse tissue sections presents specific technical challenges. For example, CD4 and CD8 have been difficult to detect using IHC on formalin-fixed and paraffin-embedded mouse tissue, prompting alternative methods. We investigated the use of formalin-free zinc-salt fixation (ZN) and optimized IHC protocols for detecting a panel of immune cell-related markers (CD3, CD4, CD8, Foxp3, B220, F4/80, CD68, and major histocompatibility complex [MHC] class-I, MHC class-II, and Gr-1). The IHC results for these markers were compared on mouse spleen tissue treated with neutral buffered formalin (NBF) or ZN with or ZN without antigen retrieval (AR). Whereas CD4 and CD8 were not detected in NBF-treated tissue, all markers were detected in ZN-treated tissue without AR. Thus, the use of ZN treatment for IHC staining can be a good tool for studying immunoreactive lesions in tissues.

AB - Tissue localization of immune cells is critical to the study of disease processes in mouse models of human diseases. However, immunohistochemistry (IHC) for immune cell phenotyping in mouse tissue sections presents specific technical challenges. For example, CD4 and CD8 have been difficult to detect using IHC on formalin-fixed and paraffin-embedded mouse tissue, prompting alternative methods. We investigated the use of formalin-free zinc-salt fixation (ZN) and optimized IHC protocols for detecting a panel of immune cell-related markers (CD3, CD4, CD8, Foxp3, B220, F4/80, CD68, and major histocompatibility complex [MHC] class-I, MHC class-II, and Gr-1). The IHC results for these markers were compared on mouse spleen tissue treated with neutral buffered formalin (NBF) or ZN with or ZN without antigen retrieval (AR). Whereas CD4 and CD8 were not detected in NBF-treated tissue, all markers were detected in ZN-treated tissue without AR. Thus, the use of ZN treatment for IHC staining can be a good tool for studying immunoreactive lesions in tissues.

KW - antigen retrieval

KW - formalin fixation

KW - immune cells

KW - immunohistochemistry

KW - zinc-salt fixation

UR - http://www.scopus.com/inward/record.url?scp=84938578681&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84938578681&partnerID=8YFLogxK

U2 - 10.1177/0192623315587593

DO - 10.1177/0192623315587593

M3 - Article

VL - 43

SP - 883

EP - 889

JO - Toxicologic Pathology

JF - Toxicologic Pathology

SN - 0192-6233

IS - 6

ER -