RNAscope™ Multiomic LS Fluorescent Assay
Visualize cell phenotypes and function with single-section spatial multiomics
The RNAscope™ Multiomic LS assay enables highly sensitive and specific detection of both RNA and proteins in the same tissue section for a true multiomic spatial assay. Proven class-leading single-molecule RNAscope technology is leveraged to enable simultaneous detection of up to 6 total proteins and RNA targets with single cell resolution.
The RNAscope™ Multiomic LS assay is designed as an automated assay for the Leica Biosystems’ BOND RX Research Advanced Staining System for high throughput multiomic spatial analysis.
Optimized protease-free multiomic workflow:
- New protease-free workflow preserves antigen integrity and allows for a wide range of antibody compatibility.
- Maintains your tissue morphology for the most accurate spatial analysis.
- Compatibility with sequential workflows or the new RNAscope Multiomic Workflow for TSA-based RNA and protein detection.
- Kit also includes protease to support your ongoing projects.
Fast and flexible assay design:
- Detect any RNA transcripts. Any gene. Any species.
- Use your own primary and secondary antibodies in sequential workflows.
- Select from pre-qualified RNAscope antibodies to accelerate multiomic assay design at higher plex.
Gold-standard RNAscope detection:
- Proven performance for FFPE samples and high to low expressing target genes.
- Backed by 10K publications.
- Over 50K probes visualized.
- Same-slide H7E staining.
Protease-free for Co-Detection and Advanced Multiomic Workflows
For more advanced multiomic capability, pre-conjugated and qualified primary and secondary antibodies are available. These ensure sensitivity and specificity minimizing laborious assay optimization.
Select up to 6 RNA targets for an all-RNA assay or combine both protein and RNA targets in 3 easy steps:
Start with up to 2 of your own primary antibodies raised in mouse and rabbit and use with available conjugated secondaries.
Add up to 4 pre-conjugated RNAscope primary antibodies from available panels.
Immuno-oncology: CD4 | CD8 | FoxP3 | PanCK
Neurobiology: NeuN | GFAP | IBA-1
Add RNA markers in open channels.
| Step | RNAscope channel assignment | ||||||
|---|---|---|---|---|---|---|---|
| C1 | C2 | C3 | C4 | C5 | C6 | ||
| 1 | Select your own primary antibodies and use with conjugated secondaries | User provided primary antibody-rabbit + RNAscope anti-rabbit-C1 | User provided primary antibody-mouse + RNAscope anti-mouse-C2 | ||||
| 2 | Select conjugated primaries from panel | RNAscope Ab C3 channel | RNAscope Ab C4 channel | RNAscope Ab C5 channel | RNAscope Ab C6 channel | ||
| 3 | Select probes in any open channel | RNAscope C1 probe | RNAscope C2 probe | RNAscope C3 probe | RNAscope C4 probe | RNAscope C5 probe | RNAscope C6 probe |
Technology Overview
The RNAscope Multiomic LS Assay allows simultaneous detection of up to six RNA or protein targets within a single section.
For RNA detection, gene-specific target probes are used, each assigned to different probe channels (- C1 through C6). The lower part of the double Z probe is designed to hybridize to its specific RNA target. The upper part of the double Z probe is specific to a particular probe channel. Channel-specific amplification trees, consisting of preamplifiers and amplifiers, are built. For protein detection, antibodies with conjugated oligonucleotides, analogous to the upper part of the double Z probe, are used and channel-specific amplification trees are then built upon these oligonucleotides.
Signal detection for each channel is performed sequentially with the addition of the final HRP-conjugated oligo and deposition of tyramide linked fluorescent dyes. The combination of ACD patented signal amplification system and the sequential tyramide deposition provides a significant boost in the signal while suppressing background noise, resulting in a highly sensitive and specific assay for both RNA and protein detection.
Visualize RNAs with RNAscope Multiplex v2 detection technology. RNA-specific target probes using proprietary double ZZ design are amplified using RNAscope technology followed by fluorescent TSA detection.
Visualize proteins using ready-to use pre-conjugated primary antibodies or your own unconjugated primary antibodies together with available pre-qualified secondary antibodies. Antibodies are detected using RNAscope amplification technology followed by fluorescent TSA detection.
Fluorophore Selection
The RNAscope Multiomic LS assay supports detection of up to 6 targets simultaneously. The following are the recommended TSA fluorophores to maximize sensitivity and minimize spectral overlap. If using other TSA fluorophores, select the best combination for your imaging system.
| Tocris TSA Flurophore | Tocris Catalog number | 3-plex |
|---|---|---|
TSA Vivid Fluorophore 520 | 323271 | ✓ |
TSA Vivid Fluorophore 570 | 323272 | ✓ |
TSA Vivid Fluorophore 650 | 323273 | ✓ |
| Opal TSA Fluorophore | Akoya Biosciences Catalog number | 3-plex | 4-plex | 5- or 6-plex |
|---|---|---|---|---|
Opal 480 Reagent Pack | FP1500001KT |
|
| ✓ |
Opal 520 Reagent Pack | FP1487001KT | ✓ | ✓ | ✓ |
Opal 570 Reagent Pack | FP1488001KT | ✓ | ✓ | ✓ |
Opal 620 Reagent Pack | FP1495001KT |
|
| ✓ |
Opal 690 Reagent Pack | FP1497001KT | ✓ | ✓ | ✓ |
Opal Polaris 780 Reagent Pack Antibody Diluent/Block (needed when using Opal Polaris 780) | FP1501001KT and ARD1001EA |
| ✓ | ✓ |
Ordering Information
| Assay components | Catalog number | Number of assays |
|---|---|---|
RNAscope™ Multiomic CORE & C1-C3 Channel Kit | 323155 323460 | 60 20 |
RNAscope™ Multiomic CORE & C1-C6 Channel Kit | 323175 323465 | 60 20 |
RNAscope™ Multiomic C4 Channel Reagents | 322950 323445 | 60 20 |
RNAscope™ Multiomic C5 Channel Reagents | 322955 323450 | 60 20 |
RNAscope™ Multiomic C6 Channel Reagents | 322960 323455 | 60 20 |
| Human TIL Panel Primary Antibodies | Catalog number | Number of assays |
|---|---|---|
RNAscope™ Ab Hs CD4-C3 | 322949 | 20 |
RNAscope™ Ab Hs CD8-C4 | 322951 | 20 |
RNAscope™ Ab Hs PanCK-C5 | 322952 | 20 |
RNAscope™ Ab Hs FoxP3-C6 | 322953 | 20 |
RNAscope™ Multiomic Human TIL Antibody Panel Pack includes CD4, CD8, PanCK and FoxP3 antibodies | 323185 | 20 |
Neuro Panel Primary Antibodies Cross reacts against mouse, human, and non-human primate targets. | Catalog number | Number of assays |
|---|---|---|
RNAscope™ Ab NeuN-C3 (Coming Soon) | AB0018-C3 | 20 |
RNAscope™ Ab GFAP-C4 (Coming Soon) | AB0028-C4 | 20 |
RNAscope™ Ab IBA-1-C5 (Coming Soon) | AB0038-C5 | 20 |
RNAscope™ Multiomic Neuronal Antibody Panel Pack includes NeuN, GFAP and IBA-1 antibodies | 321780 | 20 |
| RNAscope Secondary Antibodies | Catalog number | Number of assays |
|---|---|---|
RNAscope™ anti-rabbit-C1 | 322954 | 60 |
RNAscope™ anti-mouse-C2 | 322956 | 60 |
| Control Probes | Catalog number | Number of assays |
|---|---|---|
RNAscope™ Multiomic LS 6-ples Positive Control Probe-Hs RTU mixture of 6 probes targeting human housekeeping genes in each of 6 separate channels, Hs-POLR2A-C1, Hs-PPIB-C2, Hs-UBC-C3, Hs-HPRT1-C4, Hs-ACTB-C5, Hs-TUBB-C6 | 323198 | 30 |
RNAscope™ Multiomic LS Assay Positive Control Probe - Mm RTU mixture of 6 probes targeting mouse housekeeping genes in each of 6 separate channels, Mm-POLR2A-C1, Mm-PPIB-C2, Mm-UBC-C3, Mm-HPRT1-C4, Mm-ACTB-C5, Mm-TUBB-C6 | 322988 | 30 |
RNAscope™ Multiomic LS 6-plex Negative Control Probe | 323208 | 30 |
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