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Visualise RNA biomarkers with chromogenic or fluorescent detection using RNAscope

Example of RNAScope - CRC Tissue

 

RNAscope® is a sensitive RNA in situ hybridisation (ISH) approach for the chromogenic and fluorescent detection of RNA biomarkers.

“RNAscope® employs a unique signal amplification strategy that allows for the visualisation of target RNAs as punctate dots, where each dot represents an individual RNA molecule. The key benefits of the RNAscope® technology are high sensitivity due to its signal amplification strategy, high specificity as a result of the RNAscope® probe design minimising nonspecific off-target signals, and detection and quantification of RNA with spatial and morphological context” (Anderson et al, 2016).

We perform RNAscope® ISH on the automated Leica BOND-RXplatform which allows high throughput and reliability.  We have successfully coupled chromogenic RNAscope® ISH with protein analysis via IHC in formalin-fixed paraffin embedded (FFPE) tissue sections.  This allows the cellular localisation of single transcripts, and the identification of specific cell populations and/or cells expressing certain proteins.

We are looking forward to developing this work further by working with collaborators on new projects.

RNA Scope

FAQs

What do I need to consider if providing samples/tissue as FFPE blocks or slides? 

  • Old tissue is often poorer quality than newer tissue. We recommend re-embedding in fresh wax prior to use.
  • Tissue is better preserved in the block than on the slide, for example, historically cut slides often give poorer staining. We recommend sectioning close to the staining date.
  • Optimal formalin fixation results in better IHC or multiplex staining.
  • There should only be one tissue section per slide.
  • Place tissue as centrally as possible on the slide.
  • If intending to overlay images for analysis, tissue placement and orientation must be as similar as possible across each slide.
  • Thick tissue sections give poorer quality staining. 5-micron sections are recommended.

Please see the FAQ ‘What factors affect the quality and consistency of multiplexed images?’ for further multiplex related considerations.