quantitative rna
In Situ Hybridization

Tissue-to-data in situ hybridization services including automated staining, brightfield and fluorescence whole slide imaging capabilities, quantitative cell-by-cell data to assess RNA expression, and spatial mRNA analysis capabilities.

Advanced in situ hybridization workflow

Automated Staining: Leica BONDRX

The Leica BONDRX ensures high quality results, with advanced automated staining systems leading to reproducible protocols and high throughput. The BONDRX provides superior flexibility with automated in situ hybridization, immunohistochemistry, and multiplexing capabilities (up to 4mRNA targets).

 

High Resolution RNA Detection

Reveal’s automated in situ hybridization workflow supports chromogenic and fluorescent detection for single plex, duplex, and multiplex assays. We provide high resolution brightfield and fluorescent ISH imaging with unrivaled image quality in the cloud, and integrated QC tools to automatically assess images for focus & artifacts at scale.

 

Quantitative Cell-by-Cell Data

Reveal combines the sensitivity of the RNAscope™ assay with the imageDx™ digital pathology platform to provide quantitative data with advanced data visualization tools. Quantify RNA expression with automated cell-by-cell spot counts. Explore spatial RNA insights with co-localization data from multiplex ISH assays. Predict treatment outcomes and genomic status using AI-powered ISH models.

Certified RNAscope® Service Provider

Our scientists have undergone extensive training 
to bring you highly sensitive mRNA detection in cells and tissue through RNAscope™ technology. As a Certified RNAscope™ Service Provider, our team delivers
industry-leading data, quality, and expertise.

  • Number of ZZ Pair per target: Standard probe design is 20 ZZ probes (minimum of 6 ZZ probes)
  • Target: mRNA >300 bases, lncRNA > 300 bases
  • Applications: Data visualization for targets with spatial context when good antibodies are not available
  • Detection: Chromogenic or fluorescent, single and multiplex
  • Number of ZZ Pair per target: 1 to 3 ZZ probes based on the application
  • Target: Exon junctions/Splice Variants
    RNA 50 to 300 bases
    Validated point mutations
  • Applications: Exon junctions/splice variants, circular RNA, gene fusion, gene knockout
    Short/highly homologous sequences, TCRs and CDR sequence for T cell clones, pre-miRNA, gene editing/CRISPR, CAR-T cell validation and detection
    Point mutation, short InDel, homologues
  • Detection: Chromogenic, single to duplex
  • Number of ZZ Pair per target: N/A
  • Target: Small RNAs 17-50 bases
    ASOs, miRNAs, siRNAs
  • Applications: Small RNAs 17-50 bases
    ASOs, miRNAs, piRNAs, shRNAs, siRNAs, and tRNAs
  • Detection: Chromogenic, singleplex

powerful data insights

Assess RNA Expression

Generate data from tissue sections, regions of interest, or tissue microarrays. Data outputs include binned positive cell counts and density values. In accordance with official ACD digital analysis guidelines, positive cells are grouped into five bins (1 dot, 2-4 dots, 3-5 dots, 5-9 dots, 10+ dots) corresponding to the number of positive ISH dots per cell. Co-localization data is reported for multiplex assays.

Figure 1. Heatmap visualization of binned ISH density values across a sample set.

Simultaneous RNA & Protein Detection

Assessing RNA and protein simultaneously provides powerful information when trying to understand protein degradation or gaining insights into the regulation of expression in a sample. ISH and IHC are powerful tools that share the ability to analyze a marker at the single cell level while preserving morphological context. Explore dual ISH-IHC protocols that combine both assays on the same sample slide.

 

Figure 2. Human lung cancer with human PPIB-positive control mRNA probe in red, CAM5.2 acidic cytokeratin immunohistochemical staining in green, and DAPI counterstain. Image provided with courtesy from Advanced Cell Diagnostics, a Bio-Techne company.

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