Roche HCRTM RNA-ISH Discovery Ultra Instructions

Roche HCRTM RNA-ISH Discovery Ultra

Specifications:

• Product Name: HCRTM RNA-ISH by Molecular Instruments
• Platform: HCRTM Platform on the DISCOVERY ULTRA Platform
• Probe Type: RNA probes
• Microscopy Compatibility: Brightfield and fluorescent microscopy
• Sample Type: FFPE tissue
• Probe Design: Custom probes available at no charge
• Enzyme-free and isothermal reaction

Product Usage Instructions

How does the HCRTM RNA-ISH Platform Work?
HCRTM Amplifiers are engineered oligonucleotides that enable signal amplification by reacting with initiators to create a polymer of reporters attached to the target RNA. The platform is enzyme-free, isothermal, and compatible with both brightfield and fluorescent microscopy.

HCRTM RNA-ISH Workflow:

1. Prepare the sample using the provided HCRTM RNA-ISH Kit reagents.
2. Apply the custom probe to target the RNA sequence in the FFPE tissue.
3. Perform image analysis using either brightfield or fluorescent microscopy.

Capabilities of the HCRTM Platform on the Ventana DISCOVERY ULTRA:

• Protease-free sample preparation preserving tissue morphology
• Native compatibility for IHC integration
• Best-in-class RNA-ISH for various sample types
• Automation friendly with setup guides available
• Straightforward pricing with all necessary reagents included

Ventana DISCOVERY Chromogens Offer Key Advantages:
Translucent chromogens enable multiplexed IHC co-localization,avoiding overlapping expression issues with opaque colors like DAB and Silver.

Get to Know HCR™ RNA-ISH by Molecular Instruments

Molecular Instruments® (MI) designs and manufactures kits for automated and manual in situ hybridization (ISH) assays, offering no-charge custom probe design and compatibility with both brightfield and fluorescent microscopy.
Researchers can take advantage of the next-generation HCR™ Platform, featuring unparalleled performance, speed, and versatility. HCR™ RNA-ISH is the only assay available that has been optimized without requiring destructive enzyme/protease digestion, enabling tissue morphology preservation, native compatibility with existing IHC/IF assays, and straightforward image analysis.
MI develops and synthesizes kits powered by its innovative HCR™ Platform for bioimaging applications in academic research, drug development, and clinical pathology and diagnostics. The HCR™ Platform introduced the concept of conditional nucleic acid self-assembly – a foundational principle for the field of dynamic nucleic acid nanotechnology.

Automation on the DISCOVERY ULTRA Platform

Get to know HCR™ RNA-ISH by Molecular

Instruments!

• Achieve greater consistency from slideto-slide, day-to-day and site-to-site with automated protocols.
• Free up lab personnel – Do more valuable work, while reducing staining failure due to human error.
• Run different protocols simultaneously
  • Different techniques such as fluorescent or chromogenic IHC, FISH/CISH, and IHC & ISH on the same slide, can be run simultaneously.

How does the HCR™ RNA-ISH Platform Work?
HCR™ Amplifiers are engineered oligonucleotides that in their normal state are folded into “hairpin” shapes as depicted in the following graphic. The cascading self-assembly that results from the reaction between the initiator and the hairpins creates a polymer of reporters attached to each target thereby enabling signal amplification.
The HCR™ Platform is completely enzyme-free, isothermal at room temperature, and compatible with both fluorescent or brightfield microscopy.

Target any RNA sequence in any FFPE tissue with unparalleled performance, speed, and versatility.
This assay has been optimized without requiring destructive protease digestion, enabling tissue morphology preservation, native compatibility with existing IHC workflows, and straightforward image analysis.
Molecular Instruments leads the industry by offering complimentary custom probe design and its catalog of tens of thousands of ready-toship HCR™ Probes. Flexible probe design can also address short targets with a three week turnaround time for custom HCR™ Probes.

What are the Capabilities of the HCR™ Platform on the Ventana DISCOVERY ULTRA?

Ventana DISCOVERY Chromogens Offer Key Advantages

The Research and Development Team at Ventana have developed several new chromogens to expand ISH and IHC multiplexing capabilities in brightfield microscopy. These new chromogens are based on fluorophores to allow for unique color generation and very narrow-range light absorption to improve compatibility for ISH and IHC multiplexing. By basing IHC/ISH chromogen chemistry on fluorophores, Ventana researchers have generated new, narrow- absorbance colors such as DISCOVERY HRP-driven Purple, Red, Yellow, Blue, Green and Teal. Tyramide chemistry is used to covalently deposit these unique chromogens for excellent stability.

• Greater Control over staining intensity than traditional chromogens
• Permanently Bound HRP-driven tyramide deposition
• Distinct, strong high-quality staining
• Increases the Flexibility of multiplexing IHC and ISH
• Greater Stability in EtOH and xylene than traditional chromogens
• No Mixing! Ready-to-Use chromogens

Ready-to-Use Fluorescent Options for HCR ™ RNA-FISH

• Bright, Stable Signal covalently deposited by tyramide chemistry
• No Mixing! Ready-to-Use dispensers
• More Choices seven fluorescent options
• Widely Compatible standard emission spectra works with most imaging systems

Translucent chromogens enable multiplexed IHC co-localization.
If the biomarkers of interest are present in the same sub-cellular compartment (nucleus, cytoplasm or cell membrane) this overlapping expression of targets is a key consideration when selecting chromogen options for IHC multiplexing. When designing a multiplexed chromogenic IHC assay that includes overlapping target biomarker expression, opaque colors such as DAB and Silver should not be used.
Traditionally, analysis of overlapping targets has been accomplished using immunofluorescence, but the availability of new Ventana translucent chromogens has provided an alternative method – in brightfield! Translucent chromogens allow a color shift when both colors are present in the same cell and sub- cellular compartment.

Availability of translucent chromogens such as Purple, Yellow, and Teal has opened up the ability to visualize overlapping targets in brightfield IHC or ISH multiplexed assays.

Creating your first HCR™ RNA-ISH order is simple
Detect any RNA target with no custom design fees in three easy steps:

Ordering Information

CLICK TO ORDER

MOLECULAR INSTRUMENTS

HCR™ RNA-ISH Starter Kit

Designed to be an accessible introductory kit for evaluation of HCR™ RNA- ISH. Contains all reagents for the detection of one positive control (PPIB for human, Ppib for mouse), one negative control (dapB), one HCR™ Membrane Stain for co-detection, and HCR™ Control Slides for FFPE human/mouse liver.

| 1 kit (for 20 slides)| Varies
HCR™ Probe Kit

Contains an HCR™ Probe against any target of interest.

| 1 kit (for 20 or 90 slides)| Varies
HCR™ Detect Kit

Contains all necessary reagents for HCR™ RNA-ISH Amplification.

| 1 kit (for 20 or 90 slides)| Varies

ROCHE DIAGNOSTICS

Please contact Customer Service at 1-800-227-2155 Select Menu Option 1 for Technical Support
Select Menu Option 2 to place an order

Or contact your Roche Account Representative

For more information about
Molecular Instruments, visit our website
https://go.roche.com/molecular-instruments-starter-kits

For more about Ventana Detection, visit our website
https://go.roche.com/chromogen-resources-molecular-instruments-brochure

© 2024 Roche MC-US-13585-0124

VENTANA and DISCOVERY ULTRA are trademarks of Roche. All other product names and trademarks are the property of their respective owners.
Roche Diagnostics Corporation
9115 Hague Road
Indianapolis, Indiana 46256
diagnostics.roche.com

DISCOVERY products are for research use only, not for diagnostic purposes.

FAQ

Can I use HCRTM RNA-ISH on non-FFPE tissue samples?

The HCRTM RNA-ISH Platform is optimized for FFPE tissue samples. For non-FFPE samples, additional optimization may be required.

How long does it take to receive custom HCRTM Probes?

Custom HCRTM Probes have a turnaround time of three weeks from the order date.

Is HCRTM RNA-ISH compatible with other automation platforms?

HCRTM RNA-ISH is fully compatible with the Roche DISCOVERY ULTRA platform, with setup guides and starter kits available for automation.

References

• Roche Diagnostics
• Molecular Instruments

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