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 language | English (US) |
|---|---|
| Pages (from-to) | 883-889 |
| Number of pages | 7 |
| Journal | Toxicologic Pathology |
| Volume | 43 |
| Issue number | 6 |
| DOIs | |
| State | Published - Aug 5 2015 |
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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 journal › Article
}
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
C2 - 26157038
AN - SCOPUS:84938578681
VL - 43
SP - 883
EP - 889
JO - Toxicologic Pathology
JF - Toxicologic Pathology
SN - 0192-6233
IS - 6
ER -