Assessment of fixatives, fixation, and tissue processing on morphology and RNA integrity
Assessment of fixatives, fixation, and tissue processing on morphology and RNA integrity
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DOI：
10.1016/j.yexmp.2005.10.002
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发表时间：
2006-04-01
影响因子：
3.6
通讯作者：
Dunstan, RW
中科院分区：
文献类型：
--
作者：
Cox, ML;Schray, CL;Dunstan, RW
形态变化的分子表征需要精致的组织形态和 RNA 保存；然而，传统的固定剂通常会导致 RNA 片段化。为了优化固定组织的分子分析，我们评估了将切片固定在 70% 中性缓冲福尔马林、改良戴维森 II、70% 乙醇、UMFIX、改良 Carnoy's、改良 Methacarn、Bouin's、磷酸盐缓冲盐水或 30% 蔗糖中时大鼠肝脏的形态和 RNA 完整性。每个样品都经过标准或微波固定和标准或微波处理，并用显微镜评估切片。提取 RNA 并评估其质量和数量的保存。改良的美沙卡恩、70% 乙醇和改良的卡诺溶液各自产生的组织形态代表了福尔马林的合理替代品。修饰的 methacarn 和 UMFIX 能最好地保留 RNA 质量。相对于标准方法，微波固定和处理都不影响 RNA 完整性，尽管形态略有改善。我们得出的结论是，使用标准和微波固定和处理方法，改良的methacarn、70%乙醇和改良的Camoy溶液提供了可接受的组织形态和RNA质量的保存。在这三种固定剂中，改良的 Methacarn 提供了最好的结果，并且可以被认为是在同一样本中评估组织形态和 RNA 完整性的固定剂的选择。 (c) 2005 Elsevier Inc. 保留所有权利。
Molecular characterization of morphologic change requires exquisite tissue morphology and RNA preservation; however, traditional fixatives usually result in fragmented RNA. To optimize molecular analyses on fixed tissues, we assessed morphologic and RNA integrity in rat liver when sections were fixed in 70% neutral-buffered formalin, modified Davidson's II, 70% ethanol, UMFIX, modified Carnoy's, modified methacarn, Bouin's, phosphate-buffered saline, or 30% sucrose. Each sample was subjected to standard or microwave fixation and standard or microwave processing, and sections were evaluated microscopically. RNA was extracted and assessed for preservation of quality and quantity. Modified methacarn, 70% ethanol, and modified Carnoy's solution each resulted in tissue morphology representing a reasonable alternative to formalin. Modified methacarn and UMFIX best preserved RNA quality. Neither microwave fixation nor processing affected RNA integrity relative to standard methods, although morphology was modestly improved. We conclude that modified methacarn, 70% ethanol, and modified Camoy's solution provided acceptable preservation of tissue morphology and RNA quality using both standard and microwave fixation and processing methods. Of these three fixatives, modified methacarn provided the best results and can be considered a fixative of choice where tissue morphology and RNA integrity are being assessed in the same specimens. (c) 2005 Elsevier Inc. All rights reserved.