Sørdal Øystein, Qvigstad Gunnar, Nordrum Ivar Skjåk, Gustafsson Bjørn, Waldum Helge L
Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology (NTNU), Trondheim, Norway.
Appl Immunohistochem Mol Morphol. 2013 Mar;21(2):185-9. doi: 10.1097/PAI.0b013e31825a0048.
In situ hybridization (ISH) is a method that detects and localizes DNA or RNA in morphologically preserved tissue and cell preparations. The method is based on the principle that DNA or RNA will undergo hydrogen binding to complimentary sequences. Selective probes are labeled and used in order to detect specific sequences in tissues or cell preparations. Even though the method has improved over the past decades, there are still issues with sensitivity and specificity. The protocols are nonstandardized, and often time consuming due to multiple steps. In this paper, we have used a new and commercially available ISH kit for the detection of mRNA in formalin-fixed paraffin-embedded tissue. We have used both human and Mongolian gerbil tissue, and we evaluated mRNA expression of the neuroendocrine markers chromogranin A and histidine decarboxylase in both normal tissue and poorly differentiated tumor. In our experience, this method offers excellent sensitivity and specificity. The protocol is more standardized, and our results have been consistent. It is also less time consuming than conventional ISH protocols.
原位杂交（ISH）是一种在形态学保存完好的组织和细胞标本中检测并定位DNA或RNA的方法。该方法基于DNA或RNA会与互补序列发生氢键结合的原理。为检测组织或细胞标本中的特定序列，需标记并使用选择性探针。尽管在过去几十年中该方法有所改进，但在灵敏度和特异性方面仍存在问题。实验方案未标准化，且由于步骤繁多常常耗时较长。在本文中，我们使用了一种新型的市售ISH试剂盒来检测福尔马林固定石蜡包埋组织中的mRNA。我们同时使用了人类和蒙古沙鼠组织，并评估了正常组织和低分化肿瘤中神经内分泌标志物嗜铬粒蛋白A和组氨酸脱羧酶的mRNA表达。根据我们的经验，该方法具有出色的灵敏度和特异性。实验方案更标准化，我们的结果也具有一致性。而且它比传统ISH实验方案耗时更少。
