Chromogen Detection of microRNA in Frozen Clinical Tissue Samples Using LNA™ Probe Technology
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.
Key wordsChromogenic ISH Cryostat sections Frozen tissue In situ hybridization Locked nucleic acid microRNA
- 19.Nielsen BS, Holmstrom K (2013) Combined microRNA in situ hybridization and immunohistochemical detection of protein markers. In: Moll F, Colombo R (eds) Target identification and validation in drug discovery, vol 986, Methods in molecular biology. Humana Press, New York, pp 353–365CrossRefGoogle Scholar