Factors affecting immunoreactivity in long-term storage of formalin-fixed paraffin-embedded tissue sections
Factors affecting immunoreactivity in long-term storage of formalin-fixed paraffin-embedded tissue sections
复制标题
DOI：
10.1007/s00418-015-1316-4
复制
发表时间：
2015-07-01
影响因子：
2.3
通讯作者：
Mastracci, Luca
中科院分区：
文献类型：
--
作者：
Grillo, Federica;Pigozzi, Simona;Mastracci, Luca
用于免疫组织化学的福尔马林固定石蜡包埋(FFPE)组织切片中的抗原衰变是一种众所周知的现象，它可能会对翻译和研究产生影响，而保存时间的长短似乎是至关重要的。这项研究的目的是用14种常规使用的抗体评估所有可能导致人类组织预期标准化收集中的抗原腐烂的因素。FFPE对照组织连续切片采用不同的保存方法(常温保存、石蜡涂膜(A(R)保护)、石蜡包埋和4A℃冷藏)，保存时间分别为1、6、9、12、24和36个月。在每次截断时，用14种抗体同时对储存切片和新鲜切片进行免疫组织化学检测。将免疫反应与零时的免疫反应进行比较。部分抗体(CD3、CD31、CD117、雌孕激素受体、Ki67、P53、TTF-1、波形蛋白)免疫染色减弱，而其他抗体(平滑肌肌动蛋白、角蛋白7、20、AE1/AE3、34βE12)未见抗原衰退。抗原性的丧失与组织切片年龄成正比，并取决于储存方式，其中冷藏载玻片受影响最小。所有免疫信号减弱的抗原都是核或膜抗原，都需要进行热预处理才能修复抗原。与其他研究结果不同的是，当分析前因素被严格控制和标准化时，抗原衰变似乎仅限于需要热抗原回收的核或膜抗原。
Antigen decay in archival formalin-fixed paraffin-embedded (FFPE) tissue sections for immunohistochemistry is a well-known phenomenon which may have repercussions on translational and research studies and length of storage time appears fundamental. The aim of this study was to evaluate all possible factors which may lead to antigen decay on a prospective standardized collection of human tissues with a panel of 14 routinely used antibodies. Serial slide sections from FFPE control tissues were stored using different methods (routine storage at room temperature, Parafilm(A (R)) protected, paraffin coated and cold stored at 4 A degrees C) and for different time periods: 1, 6, 9, 12, 24 and 36 months. Immunohistochemistry was performed at each time cutoff simultaneously on stored sections and on freshly cut sections using a panel of 14 antibodies. Immunoreactivity was compared with immunoreactions performed at time zero. Reduction in immunostaining was observed for a subset of antibodies (CD3, CD 31, CD117, estrogen and progesterone receptors, Ki67, p53, TTF-1, vimentin) while for others (smooth muscle actin, keratins 7, 20, AE1/AE3, 34 beta E12), no antigen decay was observed. Loss of antigenicity was proportional to tissue section age and was dependent on mode of storage with cold storage slides being the least affected. All antigens with reductions in immunosignal were nuclear or membranous, and they all required heat pre-treatment for antigen retrieval. In contrast to results from other studies, when pre-analytical factors are strictly controlled and standardized, antigen decay seems to be restricted to nuclear or membrane antigens which require heat antigen retrieval.