A novel semi-automated in situ hybridisation protocol for microRNA detection in paraffin embedded tissue sections.
MicroRNAs (miRNAs) are small non-coding RNAs that modulate gene expression by binding to complementary sequences on target messenger RNA transcripts. In situ hybridisation (ISH) methods have been applied to the study of miRNA in tissue samples in order to understand which is the source of the miRNA of interest. In this paper, the authors describe a novel semi-automated bright field ISH method to visualise miRNAs in formalin fixed paraffin embedded tissue sections. The relevance of this work resides in the use of 3,3'-diaminobenzidine and peroxidase as the detection method, which provides a good defined deposition within tissues This method, which reveals the cells of origin of specific miRNAs, will enable investigators to further explore the biological role of miRNAs.
Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS, Istituto Nazionale Tumori, Milano, Italy.
Ambra Vittoria
AV
Gualeni
Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS, Istituto Nazionale Tumori, Milano, Italy.
Chiara Costanza
CC
Volpi
Department of Pathology, Centro di Riferimento Oncologico Aviano, IRCCS, Istituto Nazionale Tumori, Aviano, Italy.
Antonino
A
Carbone
Department of Diagnostic Pathology and Laboratory Medicine, Fondazione IRCCS, Istituto Nazionale Tumori, Milano, Italy.
Annunziata
A
Gloghini
D057205:Automation, Laboratory; D001555:Benzene Derivatives; D001706:Biopsy; D045744:Cell Line, Tumor; D004076:Digoxigenin; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D035683:MicroRNAs / Q000235:genetics*; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology; D016612:Paraffin Embedding; D009195:Peroxidase; D011237:Predictive Value of Tests; D016707:Tissue Fixation; D057188:Workflow
C557039:3,3'-diaminobenzidene; D001555:Benzene Derivatives; D005404:Fixatives; D035683:MicroRNAs; D005557:Formaldehyde; D009195:Peroxidase; D004076:Digoxigenin
IN SITU HYBRIDISATION; LYMPH NODES; LYMPHOMA; MOLECULAR PATHOLOGY