‌RNAScope IN SITU HYBRIDIZATION

Single cell, spatial gene expression for any target in any tissue

Robust and sensitive for rapid quantification of any RNA target – whilst maintaining spatial integrity

RNAscope™ is a powerful multiplex fluorescent and chromogenic in situ hybridization technology, enabling detection and quantification of target RNA within intact cells.

With its proprietary probe design, the technology enables simultaneous amplification of target-specific signals and suppression of background noise from non-specific hybridization.

RNAscope, complements microarray and PCR by retaining clinically relevant cellular and tissue context, as well as spatial variation of expression patterns.

Four channel extract from a 12-plex RNAscope HiPlex V2 assay on intestinal crypts produced at Propath
GFP detection in glomeruli by RNAscope stained at Propath on the Roche Ventana Discovery Ultra using the Discovery mRNA Purple HRP Detection kit

Propath: An experienced service provider for RNAscope analysis

A complete managed service for RNAscope, BaseScope™ in situ hybridization: from study design and sample prep through to quantitative data analysis.

Simply send us your samples and receive publication ready data in as little as four weeks.

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  • Experienced and helpful staff
    PhD-level scientists experienced at designing and running RNAscope and BaseScope, including multiplexed RNAscope methods
  • Dual ISH-IHC staining
    Expertise in combining both technologies in one single protocol on a single section
  • Automated staining
    Ability to run RNAscope on the Leica Bond or the Roche Ventana automated platforms, providing more consistent and reproducible outcomes
  • ‌Robust QA
    All processes at Propath are underpinned by well developed QA, data integrity and training protocols
  • Quantitative analysis
    An end-to-end service, which includes quantitative image analysis using the versatile Visopharm analysis platform
  • Orthogonal validation
    Access to a range of platforms for spatial biology in house, we regularly perform orthogonal multi-omic analysis within the same study
  • Rapid turnaround
    Studies can be designed and run within as little as 4 to 6 weeks, including production of publication-ready quantitative data
  • ‌100% traceability of samples
    We appreciate the irreplaceable nature of many tissues and therefore ensure complete traceability of all tissue from receipt to archive

RNAscope as an alternative to IHC

RNAscope in situ hybridization is being used by researchers to complement IHC and as an effective alternative to IHC

Any Target

Detect virtually any gene, in any species, including non-coding RNA

Speed

Save precious time and costs, bypassing the frustrations of antibody screening

‌Data Quality

Superior signal to noise ratio; and, for many targets, more robust and sensitive than IHC

Features of RNAscope – at a glance

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No antibody? No problem. Detect mRNA for targets with no or poor antibodies
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Compatible with virtually any tissue
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Potential to multiplex targets; or combine RNAscope with IHC in a single protocol on a single slide
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Featured in over 7,900 publications across a range of research areas
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Broad catalogue of over 45,000 targets in over 140 species
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Overcomes lack of manufacturing standardisation and poor reproducibility of some antibodies
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Compared with microarray and PCR, retains clinically relevant information regarding cellular and tissue context
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Single-molecule detection sensitivity enables targeting of low-expressing genes and even non-coding RNA

Propath’s workflow for the RNAscope™ In Situ Hybridization Assay

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Study design

‌We work in consultation with you to create an effective study design, considering your RNA targets and scientific objectives. We prepare a study plan for your approval and provide regular updates through the course of the study.

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Your samples

‌We handle them as though they are precious - with 100% traceability through our facility.

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‌Permeabilise

Tissue sections or cells are fixed onto slides and pretreated with RNAscope™ pretreatment reagents to unmask target RNA and permeabilize cells.

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Hybridize

Designed with ~20 target-specific double Z probes, RNAscope™ probes hybridize to target RNA molecules.

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Amplify

We apply detection reagents to amplify the hybridisation signals via sequential hybridisation of amplifiers and label probes.

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Visualize

Each punctate dot signal represents a single test target RNA molecule and can be visualized with a microscope.

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Quantify

We quantify single-molecule signals on a cell-by-cell basis by manual counting or automated image analysis using Visiopharm Software.  Alternatively we can provide high quality scans for you to undertake your own analysis.

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Tell us about your study

We’ll explain the complete process and provide as much support as needed to help you make key decisions and generate meaningful data that achieves your objectives.

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Select your panel

Choose from one of NanoString validated protein or mRNA panels, with the option to spike-in additional targets. Alternatively, we can help you design your own custom panel.

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Send us your samples

As little as 30ng of material per sample is sufficient. We treat all samples as though they are precious, with 100% traceability throughout our facility.

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Sample preparation

We perform all sample prep, including RNA extraction and thorough QC checks. We only proceed if samples meet QC requirements. We can also take sections from FFPE samples to enable interpretation of data in the context of histopathology.

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Hybridisation of RNA with molecular barcodes

RNA is hybridised with NanoString’s proprietary molecular probes. Once hybridised, the probes are immobilised and aligned on cartridges ready for counting.

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Digital counting

Hybridised molecular barcodes are then digitally counted using NanoString’s nCounter platform. Counts are precise and highly reproducible.

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Data analysis

We perform comprehensive data analysis, with outputs tailored to your specific requirements. Or we send back data files for you to perform your own analysis.

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QA review and audits

We perform comprehensive data analysis, with outputs tailored to your specific requirements. Or we send back data files for you to perform your own analysis.

RNAscope’s unique signal amplification strategy

In order to substantially improve the signal-to-noise ratio of RNA ISH, RNAscope requires two independent probes (double Z probes) to hybridize to the target sequence in tandem in order for signal amplification to occur.

As it is highly unlikely that two independent probes will hybridize to a non-specific target right next to each other, this design concept ensures selective amplification of target-specific signals.

RNAscope enhances the signal-to-noise ratio by requiring two independent probes to hybridize to the target sequence.

Advanced platforms for spatial biology available today at Propath

‌Our experience across a range of platforms for spatial biology allows us to recommend the most appropriate platform for each research project.

We understand the strengths and limitations of each platform – and how to get the best out of them.

‌NanoString GeoMx and CosMx
‌Whole slide spatial analysis of 18,000 RNA transcripts (GeoMx) and 1,000 RNA transcripts at sub cellular level (CosMx)
‌Validated for GCP studies
‌Lunaphore COMET
Highly multiplexed sequential immunofluorescence for up to 40 protein on a single slide
‌Validated for GCP studies
‌Akoya PhenoCycler and PhenoImager
‌Ultra-high plex single-cell spatial phenotyping; and focused mIF panels for high-throughput translational research
‌Validated for GCP studies
‌RNAScope and BaseScope
Robust in situ hybridization technology for rapid quantification of any RNA target in any tissue with spatial context

Outsourced R&D - Done better.

At Propath we approach biopharma R&D differently. Say goodbye to Large CRO frustrations –and hello to a more responsive, agile and efficient level of service.