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.
authors.affiliation: Center of Comparative Medicine, University of California, Davis, California, USA.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Mori
authors.affiliation: Center of Comparative Medicine, University of California, Davis, California, USA.
authors.forename: Pan
authors.initials: P
authors.lastname: Soonsawad
authors.affiliation: Center of Comparative Medicine, University of California, Davis, California, USA.
authors.forename: Louis
authors.initials: L
authors.lastname: Schuetter
authors.affiliation: Center of Comparative Medicine, University of California, Davis, California, USA.
authors.forename: Qian
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Center of Comparative Medicine, University of California, Davis, California, USA.
authors.forename: Neil E
authors.initials: NE
authors.lastname: Hubbard
authors.affiliation: Center of Comparative Medicine, University of California, Davis, California, USA Department of Pathology and Laboratory Medicine, School of Medicine, University of California, Davis, California, USA.
authors.forename: Robert D
authors.initials: RD
authors.lastname: Cardiff
authors.affiliation: Center of Comparative Medicine, University of California, Davis, California, USA Department of Pathology and Laboratory Medicine, School of Medicine, University of California, Davis, California, USA adborowsky@ucdavis.edu hmori@ucdavis.edu.
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Borowsky
mesh_terms: D000818:Animals; D000941:Antigens / Q000032:analysis; D015415:Biomarkers / Q000032:analysis*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D005404:Fixatives; D005557:Formaldehyde; D007111:Immunity, Cellular / Q000187:drug effects*; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D013154:Spleen / Q000166:cytology / Q000276:immunology; D016707:Tissue Fixation / Q000379:methods*; D015032:Zinc / Q000737:chemistry*
chemical_list: D000941:Antigens; D015415:Biomarkers; D005404:Fixatives; D005557:Formaldehyde; D015032:Zinc
keywords: antigen retrieval; formalin fixation; immune cells; immunohistochemistry; zinc-salt fixation.