评估基于锌盐的固定剂对保存抗原决定簇的作用，用于小鼠免疫系统细胞标志物的免疫组织化学显示。

Evaluation of zinc salt based fixatives for preserving antigenic determinants for immunohistochemical demonstration of murine immune system cell markers.

作者信息

Hicks D J, Johnson L, Mitchell S M, Gough J, Cooley W A, La Ragione R M, Spencer Y I, Wangoo A

机构信息

Departments of Pathology, Veterinary Laboratories Agency, New Haw, Addlestone, Surrey, KT15 3NB, United Kingdom.

出版信息

Biotech Histochem. 2006 Jan-Feb;81(1):23-30. doi: 10.1080/10520290600725375.

DOI:10.1080/10520290600725375

PMID:16760124

Abstract

Conventional aldehyde based fixatives produce good morphological preservation. However, owing to their cross-linking mechanism of action, epitope loss may occur during fixation compromising the tissue for subsequent immunohistochemical (IHC) analysis. IHC is an important tool for characterizing antigen, cytokine and cytomorphological markers. The increasing use of mouse models for study of pathogenesis has highlighted the need to investigate alternative fixatives. In the study reported here, tissue samples from RIII mice with immune mediated lesions, Mycobacterium bovis infected mice, and uninfected control mice were fixed in either zinc salt fixative or buffered formalin, then tested for IHC using a panel of antibodies (CD3, CD4, CD8, CD45, CD54, F4/80, Interferon-gamma and MIP2). Zinc salt fixation preserved processing-sensitive murine cell markers (CD4, CD8 and CD54) and improved the intensity of immunolabeling for CD45, F4/80 and CD3. Buffered formalin failed to preserve any of the processing-sensitive murine epitopes for demonstration by subsequent IHC.

摘要

传统的基于醛类的固定剂能产生良好的形态学保存效果。然而，由于其交联作用机制，在固定过程中可能会发生表位丢失，从而影响后续免疫组织化学（IHC）分析的组织样本。免疫组织化学是表征抗原、细胞因子和细胞形态学标志物的重要工具。用于发病机制研究的小鼠模型的使用日益增加，凸显了研究替代固定剂的必要性。在本报告的研究中，将患有免疫介导性病变的RIII小鼠、感染牛分枝杆菌的小鼠和未感染的对照小鼠的组织样本用锌盐固定剂或缓冲福尔马林固定，然后使用一组抗体（CD3、CD4、CD8、CD45、CD54、F4/80、干扰素-γ和MIP2）进行免疫组织化学检测。锌盐固定保存了对处理敏感的小鼠细胞标志物（CD4、CD8和CD54），并提高了CD45、F4/80和CD3免疫标记的强度。缓冲福尔马林未能保存任何对处理敏感的小鼠表位以供后续免疫组织化学显示。