RNA in situ hybridization

Precision Cancer Pathology group provides a service for RNA in situ hybridization for formalin fixed paraffin embedded tissue sections, fixed cultured cells and fixed/frozen tissue sections. You can choose either singleplex or duplex chromogenic detection, or multiplex fluorescent detection of your gene or genes of interest. We are using RNAScope and BaseScope probes and assay reagents, which allow sensitive and specific detection and localization of a single RNA transcript or other RNA molecule in your tissue sample. So far we have used 131 different RNAScope and BaseScope probes targeting human, mouse and viral genes. A list of all of the used probes can be found below.

Singleplex chromogenic RNAScope assay can be used to detect single target RNA with either HRP-DAB or AP-FAST RED kit in FFPE fixed tissue samples. Duplex detection can be used for simultaneous visualization of two RNA targets while maintaining single cell resolution using  HRP-GREEN and AP-FAST RED.

The BaseScope assay enables the detection of exon junctions/splice variants, short/highly homologous RNA sequences (50-300 bases), and point mutations at single cell sensitivity. Here we used BaseScope assay to detect AKT2-PBX4 fusion gene expression in human ovarian carcinoma FFPE sample.

RNAScope multiplex fluorescent RNA in situ hybridization assay enables single-molecule detection of 1-4 RNA targets simultaneously within intact cells using three fluorescent channels: fluorescein, Cy3, Tx-red and Cy5. The nuclei are counter stained and visualized using DAPI fluorescent dye. Multiplex assay is suitable for FFPE or fixed/fresh frozen sections as well as for fixed adherent cells.

We also provide simultaneous detection of RNA and protein in fixed adherent cells and FFPE tissue samples. Fluorescence detection can be performed using either fluorescein, Cy3 or Cy5 channels.