Abstract
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.
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Nielsen, B.S., Møller, T., Holmstrøm, K. (2014). Chromogen Detection of microRNA in Frozen Clinical Tissue Samples Using LNA™ Probe Technology. In: Nielsen, B. (eds) In Situ Hybridization Protocols. Methods in Molecular Biology, vol 1211. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1459-3_7
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DOI: https://doi.org/10.1007/978-1-4939-1459-3_7
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1458-6
Online ISBN: 978-1-4939-1459-3
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