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RNAscope ISH Technology Automated Assays - Leica

The RNAscope in situ Hybridization Technology has been developed for both manual and automated protocols.  If you require high throughput or an automated solution, ACD has partnered with Leica Biosystems to deliver RNAscope ISH automated assays.  

RNAscope 2.5 Assays, available in Brown, Red and Multiplex, RNAscope 2.5 LS Reagent Kits, RNAscope LS Duplex Reagent Kits, BaseScope and miRNAscope Assays have been developed for the Leica automated IHC ISH Platform. 

RNAscope 2.5 Leica Assay - BROWN

Designed to provide more intense Diaminobenzidine (DAB) staining.

Ideal for low copy target gene expression (1-20 copies per cell).

Visualized with bright-field microscopy.

RNAscope 2.5 Leica Assay - RED

Offers high contrast with Fast Red dye.

Red dots are clearly distinguished against the hematoxylin staining.

Visualized with standard bright-field microscopy.

BaseScope Leica Assay - RED

Detection of short RNA targets and exon junctions.

High contrast Fast Red dye, distinct against hematoxylin staining.

Visualized with bright-field microscopy.

miRNAscope Leica Assay - RED

Detects small RNAs including the detection ASOs, miRNAs, siRNSa, and smaller RNAs (17-50 bases).

miRNAscope LS probes are not compatible with RNAscope LS or BaseScope LS reagents.

RNAscope Leica Multiplex Fluorescent Assay

Simultaneous detection of up to four RNA targets, with up to four target probes.

Four target probes, each designed a different probe channel - C1, C2, C3 or C4.

Nuclei can be counter stained and visualized using DAPI fluorescent dye.

RNAscope 2.5 Leica Duplex Assay

Designed for simultaneous in situ detection of two RNA species.

Two targets are distinguished in probe Channels 1 (C1, DAB) and Channel 2 (C2, Fast Red).

Visualized with a bright-field microscopy.

RNAscope 2.5 LSx Reagent Kit - BROWN

Pre-filled to accommodate the processing of 60 slides.

Reagents come pre-filled in Bond Rx containers fixed on two trays; include Fast Red chromogen, hematoxylin, bluing and proteases.

Results are comparable to the existing RNAscope 2.5 LS assay.

RNAscope 2.5 LSx Reagent Kit - RED

Pre-filled to accommodate the processing of 60 slides.

Reagents come pre-filled in Bond Rx containers fixed on two trays; include Fast Red chromogen, hematoxylin, bluing and proteases.

Results are comparable to the existing RNAscope 2.5 LS assay.

Leica Automated Assays

 

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 

miRNAscope 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 

Leica BOND RX System 

Assay Type 

Chromogenic 

Chromogenic 

Chromogenic 

Fluorescent 

Chromogenic 

Chromogenic 

Dye Used 

Diaminobenzidine (DAB) 

Fast Red 

DAB & Fast Red Option for Green & Fast Red 

TSA-based Opal fluorophores 

Fast Red 

Fast Red 

Multiplexing 

Singleplex 

Singleplex 

Singleplex, Duplex 

Single to Fourplex 

Singleplex 

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 

Robust and specific detection of small RNAs 

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 

Detection of small RNAs including ASOs, miRNAs, piRNAs, shRNAs, siRNAs, and tRNAs 

Novel Gene or Unknown Expression 

Compatible 

Compatible 

Compatible 

Compatible 

Compatible 

Compatible 

Archival Specimens 

Compatible 

Compatible 

Compatible 

Compatible 

Compatible 

Compatible 

Microscope Imaging System 

Standard bright field 

Standard bright field 

Standard bright field 

Multispectral fluorescent imaging 

Standard bright field 

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 

FFPE  Fixed Frozen  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 

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) 

60 (20 mm x 20 mm) 

Target Sequence length required 

300-1000bp 

300-1000bp 

300-1000bp 

300-1000bp 

50-300bp 

17-50bp