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Best-in-class RNA ISH for your automated system of choice

For labs that require high throughput and automated RNA in situ hybridization (ISH) analysis, ACD has partnered with Leica Biosystems and Roche Tissue Diagnostics to deliver RNAscope® ISH automated assays.

RNAscope® 2.5 LS Reagent Kits and RNAscope® LS Duplex Reagent Kits are offered for a fully automated walk-away solution on the Leica BOND RX System by Leica Biosystems automating the Permeabilize, Hybridize and Amplify steps of the RNAscope® Workflow.

RNAscope® 2.5 VS Reagent Kits are offered for an automated walk-away solution on the DISCOVERY ULTRA and DISCOVERY XT automated tissue staining systems by Roche Tissue Diagnostics automating the Hybridize and Amplify steps of the RNAscope® Workflow. Most recently the RNAscope® VS Universal Assays offered exclusively for the DISCOVERY ULTRA allows for additional capability such as ISH-IHC or IF with Roche Tissue Diagnostic’s mRNA Universal protocol

RNAscope® Automated LS Assay

RNAscope® Automated VS Assay

RNAscope® In Situ Hybridization Assay Workflow

  • Step 1:

    Permeabilize

    Tissue sections or cells are fixed onto slides and pretreated with RNAscope® Pretreatment Kit to unmask target RNA and permeabilize cells.

  • Step 2:

    Hybridize

    Designed with ~20 target-specific double Z probes, RNAscope® Probes hybridize to target RNA molecules.

  • Step 3:

    Amplify

    RNAscope® Detection Reagents amplify the hybridization signals via sequential hybridization of amplifiers and label probes.

  • Step 4:

    Visualize

    Each punctate dot signal represents a single test target RNA molecule and can be visualized with a microscope.

  • Step 5:

    Quantify

    Single-molecule signals can be quantified on a cell-by-cell basis by manual counting or automated image analysis using HALO Software.

Automated
Automated

Which Automated Assay is right for me?

RNAscope® 2.5 Leica Assay - BROWN

RNAscope® 2.5 Leica Assay - RED

RNAscope® 2.5 Leica Duplex Assay

RNAscope® Leica Multiplex Fluorescent Assay

BaseScope™ Leica Assay - RED

Automated System

Leica BOND RX System

Leica BOND RX System

Leica BOND RX System

Leica BOND RX System

Leica BOND RX System

Assay Type

Chromogenic

Chromogenic

Chromogenic

Fluorescent

Chromogenic

Dye Used

Diaminobenzidine (DAB)

Fast Red

Diaminobenzidine (DAB) & Fast Red Option for Green & Fast Red

TSA-based Opal fluorophores

Fast Red

Multiplexing

Singleplex

Singleplex

Singleplex, Duplex

Single to Fourplex

Singleplex

Key Benefit

Robust, sensitive, permanent stain Most widely used RNAscope assay

Provides bright color stains with high contrast to background

Simultaneous in situ detection of two RNA species

Utilizes up to 4 different spectral channels providing high flexibility

Specific detection of splice variants/exon junctions and short targets

Ideal for

High throughput Routine applications

Studies of tissues with endogenous color background such as melanin in skin, liver, or lung from smokers

Co-localization studies to map co-expression of two targets (e.g. secreted ligand and its receptor)

Co-localization studies of up to 4 gene simultaneously Experimental application requiring flexibility

Detection of splice variants and exon junctions, short insertions/deletions, short targets of complex & highly homologous gene families

Novel Gene or Unknown Expression

Compatible

Compatible

Compatible

Compatible

Compatible

Archival Specimens

Compatible

Compatible

Compatible

Compatible

Compatible

Microscope Imaging System

Standard bright field

Standard bright field

Standard bright field

Multispectral fluorescent imaging

Standard bright field

Sample Type

FFPE
Fixed Frozen Tissues
Fresh Frozen Tissues
Cultured cells

FFPE
Fixed Frozen Tissues
Fresh Frozen Tissues
Cultured cells

FFPE
Fixed Frozen Tissues
Fresh Frozen Tissues
Cultured cells

FFPE Tissues(TMAs)
Fixed Frozen Tissues
Fresh Frozen Tissues
Cultured cells

FFPE
Fresh Frozen Tissues
Cultured cells

Quantitation Method

Visual inspection
HALO® SOFTWARE
Any open-source image analysis software

Visual inspection
HALO® SOFTWARE
Any open-source image analysis software

Visual inspection
HALO® SOFTWARE
Any open-source image analysis software

Visual inspection
HALO® SOFTWARE
Any open-source image analysis software

Visual inspection
HALO® SOFTWARE
Any open-source image analysis software

# of tissue sections per kit

60 (20 mm x 20 mm)

60 (20 mm x 20 mm)

60 (20 mm x 20 mm)

60 (20 mm x 20 mm)

60 (20 mm x 20 mm)

Target Sequence length required

300-1000bp

300-1000bp

300-1000bp

300-1000bp

50-300bp

X
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