Multiplexed miRNA Fluorescence In Situ Hybridization for Formalin-Fixed Paraffin-Embedded Tissues
Multiplexed miRNA fluorescence in situ hybridization (miRNA FISH) is an advanced method for visualizing differentially expressed miRNAs, together with other reference RNAs, in archival tissues. Some miRNAs are excellent disease biomarkers due to their abundance and cell-type specificity. However, these short RNA molecules are difficult to visualize due to loss by diffusion, probe mishybridization, and signal detection and signal amplification issues. Here, we describe a reliable and adjustable method for visualizing and normalizing miRNA signals in formalin-fixed paraffin-embedded (FFPE) tissue sections.
Key wordsmiRNA Fluorescence in situ hybridization Formalin-fixed and paraffin-embedded tissues Molecular diagnostics Multiplexing Signal amplification methods
N.R. is supported through a K08 award (NS072235) from the National Institute of Neurological Disorders and Stroke. T.T. is an HHMI investigator and supported through R01 funding from NIH CA159227 and MH080442 and a grant by the Starr Cancer Consortium. The project described was supported through the Rockefeller University Bridges to Better Medicine Technology Innovation Fund. The project was also partially supported by Grant Award Number (UL1RR024143) from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH) and NIH Roadmap for Medical Research, and its contents are solely the responsibility of the authors and do not necessarily represent the official view of NCRR or NIH.