RNAscope® Fluorescent Multiplex Assays
Spatial analysis of gene expression is an essential tool for comprehensive studies of complex, highly heterogeneous tissues, such as the brain, tumors, and developing organs. Traditional RNA-ISH can be challenging due to low sensitivity, inconsistent detection accompanied with limited plexing. RNAscope® Fluorescent Multiplex assays provide exceptional sensitivity, allowing single-molecule detection of RNA targets at the single cell level. The RNAscope® Multiplex Fluorescent assays are ideal for simultaneous detection studies of any genes in nearly any tissue-type using fluorescent labels. We offer with various levels of plexing to:
- FIND cells of interest by using cell specific markers
- CONFIRM cells by simultaneous detection of additional targets
- CHARACTERIZE cellular heterogeneity of complex tissues by mapping and identifying multiple cell subtypes simultaneously
“Adding a histological context to scRNAseq data is important for fully understanding the biology of distinct cell populations, and HiPlex RNAscope is a great way to do this. The ability to stain a single section of tissue with up to 12 mRNA probes makes HiPlex RNAscope a powerful tool for distinguishing cells types in situ. While the HiPlex RNAscope protocol is relatively straightforward to conduct, ACD Bio has always been there to help when I need sales or technical assistance. I would recommend HiPlex RNAscope to anyone looking to visualize distinct populations of cells from scRNAseq data in situ.”
-Jocob Swanson, Hospital for special surgery, NY
Which RNAscope® Multiplex kit is best for me - three kits are available
Use the table below to learn more about which RNA in situ hybridization product is right for your research needs.
RNAscope® HiPlex v2 | RNAscope® Multiplex Fluorescent v2 | RNAscope® Fluorescent Multiplex | |
|---|---|---|---|
Plexing Capability | 12 Plex | 4 Plex | 3 Plex |
Reagent Kit | 324400 RNAscope HiPlex12 Reagents Kit (488, 550, 650, 750) v2 | 323100 RNAscope Multiplex Fluorescent Reagent Kit v2 | 320850 RNAscope Fluorescent Multiplex Reagent Kit |
Area of research | Neuroscience, Oncology, Immuno-Oncology, Immunology | Neuroscience, Oncology, Immuno-oncology (CD8, PD1, PDL1) | Neuroscience |
Tissue Compatibility | FFPE, Fresh Frozen, Fixed Frozen | FFPE, Fixed Frozen (low expressors)** Cells and Fresh Frozen (if highly autofluorescent) | Fresh Frozen, Cells, Fixed Frozen (high expressors) |
Recommended for | High and low autofluorescing tissues | High autofluorescing tissues | Low autofluorescing tissues |
Assay TAT | 9 hours | 14 hours | 6-8 hours |
Cleavable Fluorophore | Yes | No | No |
ISH-IF compatibility | Compatible | Compatible: please refer to Doc# 323100-TN | Not recommended |
Probes Designation | RNAscope® HiPlex Probes: T1, T2, T3, T4, ……T12 | RNAscope® Probes: C1, C2, C3, C4 | RNAscope® Probes: C1, C2, C3 |
Additional Reagents required | No | Opal dyes must be purchased from Akoya Biosciences. | No |
Recommended Fluorophores | Alexa Fluor-488, Dylight 550, Dylight 650, Alexa Fluor-750 (These Fluorophores are provided in the Reagent Kit) | 3-plex: | Fluorophores included in the Reagent Kit - Alexa Fluor-488 Atto-550 and Atto-647N |
Recommended Filter sets on the Fluorescent microscope: | Ex Max 358 nm/Em Max 461 nm (DAPI) | Ex Max 358 nm/Em Max 461 nm (DAPI) | Ex Max 358 nm/Em Max 461 nm (DAPI) |
Find more information at | |||
Description | RNAscope® Fluorescent Multiplex Reagent Kit designed using ACD's proprietary amplification strategy. The assay uses Alexa Fluor-488, Atto-550, Atto-647N and Alexa Fluor-750 for detecting up to 12 targets on the same tissue section. | TSA-based Fluorescent RNAscope®detection assay for up to 4-plex capability using Opal fluorophores (sold separately). Ideal for FFPE tissues | RNAscope® Fluorescent Multiplex Reagent Kit employs Alexa Fluor-488, Atto-550 and Atto-647N fluorophores for up to 3-plex detection capability. |
