Evaluation of the value of frozen tissue section used as "gold standard" for immunohistochemistry
Evaluation of the value of frozen tissue section used as "gold standard" for immunohistochemistry
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DOI：
10.1309/7cxuyxt23e5al8kq
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发表时间：
2008-03-01
影响因子：
3.5
通讯作者：
Taylor, Clive R.
中科院分区：
文献类型：
--
作者：
Shi, Shan-Rong;Liu, Cheng;Taylor, Clive R.
为了研究使用丙酮或乙醇固定的冷冻组织切片作为免疫组化分析的“金标准”，我们评估了冷冻切片的各种条件下的固定和抗原修复（AR）。新鲜人体组织在OCT中冷冻。相邻组织块在10%中性缓冲福尔马林（NBF）中固定并石蜡包埋（FFPE）。通过6种方案固定冷冻切片：丙酮、乙醇、NBF（2个持续时间）和NBF +氯化钙（2个持续时间）。超过一半的抗体（16/26 [62%]）显示丙酮和NBF固定切片之间的免疫组织化学结果无法区分; 8（31%）在NBF和AR后显示更好的免疫组织化学信号; 2个在丙酮固定切片上得到更好的免疫组织化学结果。大多数细胞质蛋白（10/13）在丙酮和NBF固定切片之间显示出相当的免疫组织化学信号。对于核蛋白，NBF固定切片比丙酮固定切片具有更好的免疫组化信号。在大多数情况下，NBF产生更强的信号，背景更少，形态更好。这些数据不支持使用丙酮固定的冷冻组织切片作为免疫组化分析的金标准。在评价新抗体时，应结合使用丙酮和NBF固定的冷冻切片，尽管在实践中，FFPE组织切片可作为免疫组织化学分析中大多数抗原的标准。
To examine the use of acetone- or ethanol-fixed frozen tissue sections as the "gold standard" for immunohistochemical analysis, we evaluated frozen sections with various conditions of fixation and antigen retrieval (AR). Fresh human tissues were frozen in OCT. An adjacent tissue block was fixed in 10% neutral buffered formalin (NBF) and paraffin embedded (FFPE). Frozen sections were fixed by 6 protocols: acetone, ethanol, NBF (2 durations), and NBF + calcium chloride (2 durations). AR was used for all NBF-fixed sections.More than half of the antibodies (16/26 [62%]) showed immunohistochemical results indistinguishable between acetone- and NBF-fixed sections; 8 (31%) showed better immunohistochemical signals following NBF and AR; 2 gave-better immunohistochemical results for acetone-fixed sections. Most cytoplasmic proteins (10/13) showed comparable immunohistochemical signals between acetone- and NBF fixed sections. For nuclear proteins, NBF fixed sections gave better immunohistochemical signals than did acetone-fixed sections. In most cases, NBF yielded stronger signals with less background and better morphology. The data do not support the use of acetone-fixed frozen tissue sections as the gold standard for immunohistochemical analysis. In evaluating new antibodies, a combination of acetone- and NBF-fixed frozen sections should be used, although in practice, FFPE tissue sections may serve as the standard for most antigens for immunohistochemical analysis.