RNAscope™ HiPlex Pro for COMET™

RNAscope™ HiPlex Pro for COMET™

Unlock the potential of spatial multiomics

RNAscope™ Multiomic LS Fluorescent Assay

Visualize cell phenotypes and function with single-section spatial multiomics

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.

RNAscope Chemistry

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.

New Protein Capability

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		✓	✓

Note: For 5-plex assay, select any 5 of the recommended fluorophores.

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

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	AB0014-C3	20
RNAscope™ Ab GFAP-C4	AB0024-C4	20
RNAscope™ Ab IBA-1-C5	AB0034-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

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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RNAscope Multiomic LS Assay

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RNAscope™ Antibodies

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RNAscope™ Kits Now Include Reagents for Protease-Free Workflow

Welcome to the world of spatial multiomcs with RNAscope™. Now with the innovative protease-free workflow powered by PretreatPro™, combine high precision RNA detection with expanded protein biomarker co-detection for RNAscope assays on the RNascope Multiplex v2 assay, Roche DISCOVERY ULTRA and the Leica Biosystems BOND RX platforms.

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.

RNAscope™ Plus smRNA-RNA LS Assay

Small regulatory RNAs, including miRNAs, are instrumental in regulating human health and have been associated with a wide range of different diseases. Oligonucleotide therapies, including antisense oligonucleotides (ASOs) and RNA interference (RNAi), represent a unique class of gene therapies that have the potential to provide highly focused, long-lasting therapeutic effects by utilizing similar cellular processes.

RNAscope™ Plus smRNA-RNA Assay

Small regulatory RNAs, including miRNAs, are instrumental in regulating human health and have been associated with a wide range of different diseases. Oligonucleotide therapies, including antisense oligonucleotides (ASOs) and RNA interference (RNAi), represent a unique class of gene therapies that have the potential to provide highly focused, long-lasting therapeutic effects by utilizing similar cellular processes.

DNAscope Duplex Assay

The DNAscope assay is now discontinued.

Integrated Co-Detection

ACD’s new Co-detection Assays will allow researchers to simultaneously examine cell-type specific gene expression and identify cellular sources of secreted proteins. The new workflow will allow inclusion of wider range of antibodies to be combined with RNA ISH enabling researchers to acquire more data and conserve precious samples.

The new assay and reagents are compatible with the Integrated Co-detection workflow schematic below

miRNAscope™ Pretreatment Reagents

Pretreatment reagents are used for blocking endogenous peroxidase activity and to optimally permeabilize properly prepared samples. These reagents have been optimized to better access target RNA and reduce optimization time in multiple tissues such as formalin-fixed, paraffin-embedded (FFPE) (including archival) tissue, fresh‑frozen (FF) tissue, fixed‑frozen tissue, tissue microarray (TMA), and cell preparations.

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