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 | |
|---|---|---|
Plexing Capability | 12 Plex | 4 Plex |
Reagent Kit | 324400 RNAscope HiPlex12 Reagents Kit (488, 550, 650, 750) v2 | 323100 RNAscope Multiplex Fluorescent Reagent Kit v2 |
Area of research | Neuroscience, Oncology, Immuno-Oncology, Immunology | Neuroscience, Oncology, Immuno-oncology (CD8, PD1, PDL1) |
Tissue Compatibility | FFPE, Fresh Frozen, Fixed Frozen | FFPE, Fixed Frozen (low expressors)** Cells and Fresh Frozen (if highly autofluorescent) |
Recommended for | High and low autofluorescing tissues | High autofluorescing tissues |
Assay TAT | 9 hours | 14 hours |
Cleavable Fluorophore | Yes | No |
ISH-IF compatibility | Compatible | Compatible: please refer to Doc# 323100-TN |
Probes Designation | RNAscope HiPlex Probes: T1, T2, T3, T4, ……T12 | RNAscope Probes: C1, C2, C3, C4 |
Additional Reagents required | No | Opal dyes must be purchased from Akoya Biosciences. |
Recommended Fluorophores | Alexa Fluor-488, Dylight 550, Dylight 650, Alexa Fluor-750 (These Fluorophores are provided in the Reagent Kit) | 3-plex: |
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) |
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 |
