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Chromogen Detection of microRNA in Frozen Clinical Tissue Samples Using LNA™ Probe Technology

  • Boye Schnack NielsenEmail author
  • Trine Møller
  • Kim Holmstrøm
Part of the Methods in Molecular Biology book series (MIMB, volume 1211)

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.

Key words

Chromogenic ISH Cryostat sections Frozen tissue In situ hybridization Locked nucleic acid microRNA 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Boye Schnack Nielsen
    • 1
    Email author
  • Trine Møller
    • 1
  • Kim Holmstrøm
    • 1
  1. 1.Molecular HistologyBioneer A/SHørsholmDenmark

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