Product Overview
The RNAscope HiPlex v2 (manual) assay is an expansion of the RNAscope HiPlex assay platform, an advanced, in situ hybridization (ISH) tool to understand vital gene expression patterns at single cell resolution. The new assay generates precise gene expression data, enabling comprehensive spatial studies from 12 targets in FFPE samples and up to 48 targets in fresh and fixed frozen sample types.
Fluorescent Multiplex Detection
Fluorescent Multiplex Detection
•Detection of up to 12 RNAscope targets in FFPE tissues
•Detection of up to 48 RNAscope targets in frozen tissues with HiPlex UP
Flexible Imaging Options
Flexible Imaging Options
•Standard kit suited to systems capable of multiplex imaging with DAPI, AlexaFluor 488, Dylight 550, Dylight 650, and Alexa Fluor 750
•Alternative kit suited to systems capable of multiplex imaging with DAPI, AlexaFluor 488, Dylight 550, and Dylight 650
WSI Registration Software
WSI Registration Software
Register, merge, and background subtract images obtained using the RNAscope HiPlex v2 Assay. Whole slide images are supported with v2.1.
Technology Overview
The RNAscope HiPlex assay utilizes cleavable versions of different fluorophores and an iterative detection process that is repeated until all 12 targets are detected. The fluorophore cleaving procedure is rapid with minimal to no effect on RNA and tissue morphology since it does not strip away the bound probe. The images from various rounds can be merged using RNAscope HiPlex Image Registration Software v2.1.
Validate Gene Signatures with Spatial Context at the Single Cell Level
Visualizing striatal Drd1a and Drd2 medium spiny neurons (MSNs) using the RNAscope HiPlex assay for 12 targets in fresh frozen mouse brain sections.
Targets are Drd1a, Drd2, Foxp1, Pcdh8, Synpr, Htr7, Meis, Calb1, Crym, Cnr1, Wfs1, and TH.
TME Profiling
TME Profiling
Detect immune cells, tumor cells, chemokines, and cytokines in the tumor microenvironment (TME).
Transcriptomic Profiling
Transcriptomic Profiling
Strengthen data from single cell experiments by using RNAscope HiPlex assay for spatial mapping and confirmation of gene signatures.
Customer Success Stories
Lab of Dr Beth Stevens, Boston Children’s Hospital
RNAscope Hiplex v2 Assay Image Gallery
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Introducing the RNAscope HiPlex v2.0 assay for single-cell and spatial transcriptomic profiling
This webinar will demonstrate detection and characterization of tumor infiltrating immune cells using the new HiPlex v2 assay thereby advancing our understanding of the dynamic crosstalk within the complex and heterogeneous tumor microenvironment (TME)
Learning Objectives:
- Learn about the expanded capabilities and applications of the HiPlex v2 assay
- Understand how the assay enables characterization of immune cell types and activation states
- Study complex spatial relationships among different cell types within the TME.
RNAscope Hiplex Panel Design Guidance
| Channel | Pros | Cons | Recommendations |
|---|---|---|---|
| Fluorescein - Green For Alexa Fluor 488 | Visible with naked eye | Least distinct from most tissue autofluorescence | High Expressor |
| Cyanine 3 - Orange | Visible with naked eye | None | Low Expressor or Expression Level Unknown |
| Cyanine 5 - Far RedFor Dylight 650 | Easily differentiated from autofluorescence | Not visible to the naked eye | Low Expressor or Expression Level Unknown |
| Cyanine 7 - Far Red For Alex Fluor 750 | Easily differentiated from autofluorescence | Not visible to the naked eye | Low Expressor |
Order Checklist
| Component | Product Name | Catalog # |
|---|---|---|
Reagent Kits | 324409 | |
324419 | ||
Species Specific Positive Control Probe | 324434 | |
324311 | ||
324433 | ||
Universal Negative Probe | 324341 | |
Probe Diluent | 324301 | |
Software | 300065 | |
Hydrophobic Barrier Pen | 310018 | |
Control Slide | 310023 | |
310045 | ||
Hybridization System | HybEZ™ II Hybridization System | various |
Microscope Detection Capabilities | Fluorescent microscope with detection capability to detect DAPI, AF488, Dylight 550, Dylight 650 and AF750 | User provided |
Intro Packs
RNAscope HiPlex12 Intro Packs contain ACD consumables needed to get started using the assay, including hydrophobic barrier pen, control probes, and control slides (for human or mouse, not rat)
| Component | Name | Catalog # |
|---|---|---|
Species-specific Intro Packs with 3-color (+DAPI) capability | RNAscope Intro Pack for HiPlex12 Reagents Kit (488, 550,650,750) - Mouse | 324440 |
RNAscope Intro Pack for HiPlex12 Reagents Kit (488, 550,650,750) - Rat | 324441 | |
RNAscope Intro Pack for HiPlex12 Reagents Kit (488, 550,650,750) - Human | 324442 | |
Species-specific Intro Packs with 4-color (+DAPI) capability | RNAscope Intro Pack for HiPlex12 Reagents Kit ((488, 550,650) - Mouse | 324443 |
RNAscope Intro Pack for HiPlex12 Reagents Kit (488, 550,650) - Rat | 324444 | |
RNAscope Intro Pack for HiPlex12 Reagents Kit (488, 550,650) - Human | 324445 |