Visualize RNA with single-molecule sensitivity and single-cell resolution

Invitrogen™ ViewRNA™ ISH Cell Assays have the sensitivity and robustness to directly detect single RNA molecules at single-cell resolution and quantify gene expression. The assay employs a proprietary fluorescent in situ hybridization (FISH) and sequential branched-DNA (bDNA) amplification technique, using independent but compatible signal amplification systems to enable simultaneous detection of up to four RNA transcripts in single cells using a standard epifluorescence microscope or high-content imager.

Technology overview

ISH is a powerful technique that enables the specific detection of target RNA localization in fixed cells. Traditional FISH techniques using large oligonucleotide sequences labeled with one to five fluorophores are generally limited by high background and low sensitivity due to non-specific binding and insufficient signal amplification.

ViewRNA Cell Assays incorporate a proprietary probe set design and bDNA signal amplification technology. A target-specific Probe Set of approximately 20 oligonucleotide pairs hybridizes to the target RNA of interest. Signal amplification is achieved through specific hybridization of adjacent oligonucleotide pairs to bDNA structures, which are formed by Pre-amplifiers, Amplifiers, and fluorochrome-conjugated Label Probes. This methodology results in greater specificity, lower background, and higher signal-to-noise ratios. ViewRNA Cell Assays are the most sensitive and specific RNA ISH methods on the market.

In comparison, to create images with similar discernible spots, traditional FISH techniques can require a 600-times longer exposure and a 100-times greater camera gain than ViewRNA ISH Cell Assays. Furthermore, under equivalent imaging conditions, a ViewRNA ISH Cell Assay is 100-times brighter, creating a minimum two- to three-times higher signal-to-noise ratio (Nat Methods 10:1127 (2013) ).

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Assay kits and supporting products

The ViewRNA ISH Cell Assay includes all the assay reagents for signal amplification and detection required for visualization of one to three target RNAs (mRNA and lncRNA) using adherent or suspension cells. ViewRNA ISH Cell 740 Module is designed to be used in conjunction with ViewRNA ISH Cell Assay and allows analysis of an additional RNA target in the 740 channel. Positive control probes and target probes for genes of interest are sold separately. ViewRNA Probe Sets are designed for use with the ViewRNA ISH Cell Assays, ViewRNA Cell Plus Assay, ViewRNA ISH Tissue Assays and PrimeFlow RNA Assay.

High-throughput, single-cell quantitation and imaging of four RNA targets

Invitrogen™ ViewRNA™ High-Content Screening Assay is a 96- or 384-well plate-based assay compatible with standard liquid-handling automation systems and high-content imaging systems that can simultaneously detect four RNA targets in single cells. Unlike high-throughput PCR screening, image-based screening enables the study and subcellular localization of gene expression at the single-cell level in heterogeneous samples without the need to isolate specific cell types or the extraction, purification, and amplification of a transcript. This powerful platform can be used across the drug discovery pipeline, from primary screens of compound libraries to downstream companion assays. For more information, please contact your local account manager.

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