Evaluation of zinc salt based fixatives for preserving antigenic determinants for immunohistochemical demonstration of murine immune system cell markers
Evaluation of zinc salt based fixatives for preserving antigenic determinants for immunohistochemical demonstration of murine immune system cell markers
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DOI：
10.1080/10520290600725375
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发表时间：
2006-01-01
影响因子：
1.6
通讯作者：
Wangoo, A.
中科院分区：
文献类型：
--
作者：
Hicks, D. J.;Johnson, L.;Wangoo, A.
传统的醛基固定剂具有良好的形态保存效果。然而，由于它们的交联作用机制，表位丢失可能在固定过程中发生，影响组织随后的免疫组化（IHC）分析。免疫组化是表征抗原、细胞因子和细胞形态学标志物的重要工具。越来越多地使用小鼠模型来研究发病机制，这突出了研究替代固定剂的必要性。在这里报道的研究中，将免疫介导病变的RIII小鼠、牛分枝杆菌感染小鼠和未感染对照小鼠的组织样本固定在锌盐固定液或缓冲福尔马林中，然后使用一组抗体（CD3、CD4、CD8、CD45、CD54、F4/80、干扰素γ和MIP2）检测免疫组化。锌盐固定保存了加工敏感的小鼠细胞标记物（CD4、CD8和CD54），并提高了CD45、F4/80和CD3的免疫标记强度。缓冲福尔马林未能保存任何加工敏感的小鼠表位，以供随后的免疫组化验证。
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.