RNAscope Webinars, Videos, Presentations

Aug

28

Getting Started and Troubleshooting Tips for RNAscope^® and BaseScope™ Assays

Ann Marie Baker, PhD, Technical Support Scientist
Getting Started and Troubleshooting Tips for RNAscope^® and BaseScope™ Assays

Ann Marie Baker, PhD

Technical Support Scientist
Outline:

Outline:

The RNAscope^® and BaseScope™ Assays are based on the proprietary RNAscope^® Technology, enabling RNA target expression analysis within intact cells and tissues with high sensitivity and specificity. These represent major advancements in RNA ISH approaches.

Main features of this technology are:
- Proprietary probe design to amplify target-specific signals but not background noise from non-specific hybridization
- Capacity to provide single-cell and single-RNA molecule sensitivity in the context of complex tissue microenvironments
- Advances to this technology enabling detection of any mRNA, short RNA targets e.g. transcript variants and exon junctions
RNAscope^® and BaseScope™ Assays are ideal tools to complement technologies like IHC, qPCR or NGS. In this webinar we will discuss the benefits of both RNAscope^® and BaseScope™ assays and share tips and tricks on how to perform these assays successfully to obtain publication-quality RNA in situ results. We will also highlight some examples of data for stem cell research specific applications, such as analysis of LRG5 expression.

Who should attend:

This webinar will benefit both new and experienced RNAscope^® technology users.

Learning Objectives:

How RNAscope^® and BaseScope™ assays work

What tools are needed for getting started

Tips and tricks for sample preparation

Examples of staining results

FAQs & Troubleshooting tips

Where to find information

Examples of data featuring research applications
Dates and Registration:

August

28

09:00 am PT

Register Now

You can watch a video or download presentation of the past webinars.

Recorded Webinars

Follow a step-by-step visual guide to assist with running the RNAscope Manual Assay.

De-paraffinization

De-paraffinization is performed to ensure complete removal of the paraffin from FFPE samples to allow for the probes to penetrate the target RNA after adequate pretreatment.

Endogenous Peroxidase Blocking

RNAscope H2O2 step is performed during the RNAscope assay to block endogenous peroxidase enzyme activity to prevent hazy background after detection.Note: Pretreatment 1 refers to RNAscope H2O2 reagent and is the first pretreatment perfomed on your samples for Chromogenic assays.

Target Retrieval (Boiling)

Target Retrieval step is a heat induced epitope retrieval method that is necessary to reverse the cross-linking caused by the formalin fixation step. Note: Pretreatment 2 refers to Target Retrieval reagent.For an alternative steamer protocol refer to the appendix of User Manual Part 2 Brown and Red.

Protease Plus (Protease Digestion)

Protease Plus is a broad spectrum protease that is intended to permeabilize the samples adequately to allow the probes to reach the target mRNA.Note : Pretreament 3 refers to Protease Plus reagent.

Target Probe Hybridization

ACD provides properietary double "ZZ" oligo probes designed to hybridize to your specific RNA target

Amplification (Amp1-Amp6) with Wash Step

RNAscope detection reagents amplify hybridization signals via sequential hybridization of amplifiers.

DAB Colormetric Reaction

Chromogenic detection is based on the enzyme substrate reaction which leads to the formation of an insoluble precipitate visualized in the form of punctate dots for the RNAscope assay.

Gill's Hematoxylin Counter Stain

Hematoxylin staining is a counterstain used to provide a contrast to better visualize the signal and to observe the morphology of the sample and identify the localization of the signal.

Tissue Dehydration

The tissue dehydration step after the counterstaining step results in removal of excess moisture which provides better tissue morphology and preservation of the signal.

Mounting

The final step in RNAscope after staining requires the use of mounting media to adhere a coverslip to tissue section or cell smear. This helps protect the sample and the staining from physical damage and helps improve the clarity and contrast of an image during microscopy.

Download any of the following webinar presentation documents.

Presentations	Download File
Ready Set Go Getting Started with RNAscope_May 12 2015
Ready Set Go Getting Started with RNAscope_Apr 14 2015
RNAscope Troubleshooting Tips_June 16 2015
RNAscope Troubleshooting Tips_July 14 2015