title: Chromogen detection of microRNA in frozen clinical tissue samples using LNA™ probe technology.
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.
authors.affiliation: Molecular Histology, Bioneer A/S, Kogle Allé 2, 2970, Hørsholm, Denmark, bsn@bioneer.dk.
authors.forename: Boye Schnack
authors.initials: BS
authors.lastname: Nielsen
authors.forename: Trine
authors.initials: T
authors.lastname: Møller
authors.forename: Kim
authors.initials: K
authors.lastname: Holmstrøm
mesh_terms: D015925:Cryopreservation; D005456:Fluorescent Dyes / Q000032:analysis; D005557:Formaldehyde / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000191:economics / Q000379:methods*; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D009841:Oligonucleotides / Q000032:analysis* / Q000235:genetics; D016612:Paraffin Embedding / Q000191:economics / Q000379:methods*; D016707:Tissue Fixation / Q000191:economics / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D035683:MicroRNAs; D009841:Oligonucleotides; C477371:locked nucleic acid; D005557:Formaldehyde