Nielsen Boye Schnack, Møller Trine, Holmstrøm Kim
Molecular Histology, Bioneer A/S, Kogle Allé 2, 2970, Hørsholm, Denmark,
Methods Mol Biol. 2014;1211:77-84. doi: 10.1007/978-1-4939-1459-3_7.
Specific chromogen- and fluorescence-based detection of microRNA by in situ hybridization (ISH) in formalin-fixed and paraffin-embedded (FFPE) tissue sections has been facilitated by locked nucleic acid (LNA)-based probe technology and can be performed within a single working day. In the current method paper, we present a similar simple 1-day ISH method developed for cryostat sections obtained from clinical cryo-embedded tissue samples. The presented chromogen-based ISH method does not involve proteolytic pretreatment, which is mandatory for FFPE sections, but still retains a sensitivity level similar to that obtained in FFPE sections. The LNA-based ISH method is not only applicable in situations where only access to cryo-embedded material is possible, but it also has a potential use if combining microRNA ISH with immunohistochemistry in double fluorescence staining with antibodies not being compatible with proteolytic predigestion.
基于锁核酸（LNA）的探针技术推动了在福尔马林固定石蜡包埋（FFPE）组织切片中通过原位杂交（ISH）对微小RNA进行特定的基于色原和荧光的检测，并且可以在一个工作日内完成。在当前的方法论文中，我们展示了一种类似的简单的一日ISH方法，该方法是为从临床冷冻包埋组织样本获得的低温恒温器切片开发的。所展示的基于色原的ISH方法不涉及蛋白水解预处理（FFPE切片必需），但仍保持与FFPE切片相似的灵敏度水平。基于LNA的ISH方法不仅适用于只能获取冷冻包埋材料的情况，而且在将微小RNA ISH与免疫组织化学结合进行双荧光染色且抗体与蛋白水解酶消化不兼容时也有潜在用途。
