Comparison of Formalin-Free Tissue Fixatives A Proteomic Study Testing Their Application for Routine Pathology and Research
Comparison of Formalin-Free Tissue Fixatives A Proteomic Study Testing Their Application for Routine Pathology and Research
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DOI：
10.1043/2009-0676-oa.1
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发表时间：
2011-06-01
影响因子：
4.6
通讯作者：
Popper, Helmut H.
中科院分区：
文献类型：
--
作者：
Kothmaier, Hannelore;Rohrer, Daniela;Popper, Helmut H.
背景。-福尔马林固定，石蜡包埋的组织是世界各地病理部门实践的常规诊断处理方法。确定非交联、无福尔马林组织固定在病理学和蛋白质组学研究中的诊断价值。我们在10例患者的肺癌标本中测试了3种市售的无福尔马林组织固定剂-FineFIX、RCL 2和HOPE。使用免疫组织化学和Western印迹法评价固定剂对组织形态学、蛋白质回收率和肺癌中不同表达的选定蛋白质组的免疫反应性的影响。与标准的福尔马林固定，石蜡包埋程序相比，苏木精-伊红肿瘤细胞分析同样适用于所有测试的固定剂。Movat pentachrome染色显示了分析的不同基质和细胞蛋白的可比结果。与福尔马林固定、石蜡包埋或冷冻组织相比，RCL 2（P = .01）和HOPE固定剂（P = .03）提高了蛋白质回收率。我们的数据清楚地表明，固定剂的评价影响免疫反应匹配，福尔马林固定，石蜡包埋的肺癌组织。特别是，膜结合蛋白，如表皮生长因子受体EGFR，可以更有效地检测免疫组化和蛋白质印迹。我们已经证明，无福尔马林固定剂有可能在常规病理学和研究中取代福尔马林在组织形态学和蛋白质保存。(Arch Pathol Lab Med. 2011;135：744-752）
Context.-Formalin-fixed, paraffin-embedded tissue is the routine processing method for diagnostics practiced in pathology departments worldwide.Objective.-To determine the potential value of non-cross-linking, formalin-free tissue fixation for diagnostics in pathology and proteomic investigations.Design.-We tested 3 commercially available, formalin-free tissue fixatives-FineFIX, RCL2, and HOPE-in lung cancer specimens from 10 patients. The fixatives were evaluated for their effects on tissue morphology, protein recovery, and immunoreactivity for a selected panel of proteins differently expressed in lung cancer, using immunohistochemistry and Western blotting.Results.-Tumor-cell analysis with hematoxylin-eosin worked equally well for all tested fixatives when compared with the standard formalin-fixed, paraffin-embedded procedure. Movat pentachrome stains showed comparable results for the different matrices and cellular proteins analyzed. The RCL2 (P = .01) and HOPE fixatives (P = .03) improved protein recovery when compared with formalin-fixed, paraffin-embedded or frozen tissues. Our data clearly show that the fixatives evaluated influenced immunoreactivity to matched, formalin-fixed, paraffin-embedded lung cancer tissue. In particular, membrane-bound proteins, such as epidermal growth factor receptor EGFR, can be detected more efficiently by immunohistochemistry and Western blotting.Conclusion.-We have demonstrated that formalin-free fixatives have the potential in routine pathology and research to replace formalin in histomorphology and protein preservation. (Arch Pathol Lab Med. 2011;135:744-752)