RNA - protein co-detection with Imaging Mass Cytometry and RNAscope assay
Tumor-immune interaction in breast cancer tissue using RNAscope HiPlex with Imaging Mass Cytometry for RNA-Protein co-detection. RNAscope was used for visualizing chemokines and growth factors while antibodies were used for immune cell markers.
RNAscope™ HiPlex assay can be combined with Imaging Mass Cytometry™ (IMC™) to enable same-slide co-detection of RNA and protein targets on FFPE tissues.
IMC is a proven tool for the study of complex cellular interactions in the TME. It utilizes CyTOF® technology for simultaneous assessment of 40-plus protein markers at subcellular resolution without spectral overlap or background autofluorescence, thus providing unprecedented insight into the tissue organization and function. The RNAscope HiPlex v2 assay can detect up to 12 targets on FFPE tissues. By using metal-conjugated labeled probes instead of the Fluoros ( Fluoros T1-T4, Fluoro T5-T7, Fluori T8-T12), RNA targets can be detected using the Hyperion Imaging Mass Cytometry system.
Graphical schematic of the three-step procedure to acquire RNA and protein co-detection data with IMC
Key advantages of RNA-protein co-detection
- Spatial phenotyping with cell marker-specific antibodies and target RNA expression.
- Visualizing inflammatory signatures.
- Studying cell-type specific gene expression.
- Identifying the cellular source of secreted factors such as cytokines, chemokines and growth factors.
This technical note provides a workflow and procedure to achieve RNA and protein co-detection data in IMC experiments. This workflow utilizes the highly sensitive and specific RNAscope technology for RNA detection with the multiplexing capability of IMC to visualize key RNA and protein markers in the same FFPE samples
Flyer
RNA - protein co-detection with Imaging Mass Cytometry using the RNAscope assay
Tech Note
Co-Detection of RNA and Protein to Explore Tumor-Immune Interactions Utilizing RNAscope With Imaging Mass Cytometry
To learn more about the RNAscope™ HiPlex12 v2 Assay - please click here: https://acdbio.com/rnascope-hiplex-assays or review the user manual here
Getting started:
Step1: Here are the reagents needed for Step 1 of the workflow
ACD
Metal-ready probes (ACD, Bio-Techne) | Product Information |
|---|---|
RNAscope™ HiPlex metal-ready probes T1-T12 | 322710 |
RNAscope™ HiPlex metal-ready probe T1 | 322711 |
RNAscope™ HiPlex metal-ready probe T2 | 322712 |
RNAscope™ HiPlex metal-ready probe T3 | 322713 |
RNAscope™ HiPlex metal-ready probe T4 | 322714 |
RNAscope™ HiPlex metal-ready probe T5 | 322715 |
RNAscope™ HiPlex metal-ready probe T6 | 322716 |
RNAscope™ HiPlex metal-ready probe T7 | 322717 |
RNAscope™ HiPlex metal-ready probe T8 | 322718 |
RNAscope™ HiPlex metal-ready probe T9 | 322719 |
RNAscope™ HiPlex metal-ready probe T10 | 322720 |
RNAscope™ HiPlex metal-ready probe T11 | 322721 |
RNAscope™ HiPlex metal-ready probe T12 | 322722 |
Standard BioTools:
Labelling kit (Standard BioTools) | Product Information |
|---|---|
Maxpar X8 Antibody labeling kit | Select the metal-specific kit of your choice |
Step2: Here are the reagents needed for Step 1 of the workflow
ACD
Assay (ACD, Bio-Techne) | Product Information |
|---|---|
RNAscope™ HiPlex12 Flex Reagents Kit v2 (without Fluoros) OR RNAscope HiPlex12 Reagents Kit v2 (with Fluoros) | 322725
324409 |
Equipment (ACD, Bio-Techne) | Product Information |
|---|---|
HybEZ™ II Hybridization System | 321710 or 321720 |
HybEZ Humidity Control Tray (with lid) | 310012 |
Standard BioTools
Labelling kit (Standard BioTools) | Product Information |
|---|---|
Maxpar X8 Antibody labeling kit | Select the metal-specific kit of your choice |
Step3: Please visit the Standard BioTools website to get the suitable antibody panels.
