- Mar26How to correctly interpret your RNAscope® imagesHow to correctly interpret your RNAscope® imagesConnie Zhang, PhD.Senior Technical Support ScientistSerdar Tulu, PhD.Imaging SpecialistAmber Jolly, PhD.Image Analysis ScientistOutline:
RNAscope® assay is an innovative and proprietary RNA in situ hybridization (ISH) assay based on ACD’s patented technology with signal amplification and simultaneous background noise suppression. RNAscope® assay delivers quantitative, sensitive and specific molecular detection of RNA species on a cell-by-cell basis with anatomical context in a single assay.
Who should attend:- Pathologists who may or may not be familiar with RNAscope® ISH
- New and existing RNAscope® Assay users (manual and automated assay platforms)
- Traditional ISH users new to the RNAscope® technology
- IHC users interested in RNA ISH for complementary studies
Learning Objectives:In this webinar, we will focus on how to review and analyze RNAscope® assay data images successfully. We will cover the following topics:
• Assess examples of RNAscope® images with poor, medium and high-quality target RNA staining
• Learn best practices for accurately interpreting your RNAscope® images and discuss RNAscope® scoring tips
• Review available types of data analysis and data presentation using HALOTM and open-source software on RNAscope® images
• Learn how to best capture high quality images for subsequent data analysis
• Review the current workflow of quantitative data analysis by the Pharma Assay Services Team
Dates and Registration:
You can watch a video or download presentation of the past webinars.
- Tips and Tricks for Running the RNAscope® Multiplex Fluorescent Assay
- Tips and Tricks for Getting Started with the RNAscope® and BaseScope™ Assays
- Cancer Gene Expression Targets Validated Using Novel Histopathology Tools
- Role for NaV1.3 in the developing cerebral cortex
- Using single cell RNAseq and in situ hybridization to determine neuronal composition
- Tips and Tricks for Getting Started with RNAscope® and BaseScope™ Assays
- Application of a Novel ISH Approach to Elucidate Splice Variants in Schizophrenia
- Visualize your research breakthroughs with RNAscope® Multiplex Fluorescent assays
- The Basescope™ Duplex Product Launch Webinar - 10/30 9AM PT
- The RNAscope® Assay Data Analysis Webinar
- Getting Started and Troubleshooting Tips for RNAscope® and BaseScope™ Assays
- Harnessing in situ transcriptomics to reveal complex tissue architecture at single-cell resolution
- RNAscope® and BaseScope™ Assays are ideal tools to complement technologies like IHC, qPCR or NGS
- Mind the Transcripts: Visualizing RNA Expression and Variants within the Brain
- Elucidation of stem cell dynamics via Rspondin3 regulation using multiplex RNA in situ hybridization techniques
- Understanding heterogeneity in cancer through high-resolution imaging
- RNA in situ Hybridization: The Next Generation of Immunohistochemistry?
- BaseScope™ LS and VS Automated Assays Webinar
- Visualize your research breakthroughs with RNAscope® Multiplex Fluorescent assays
- Tips for Getting Started with RNAscope® and BaseScope™ Assays
- Webinar: Studying the Genetic Basis of Neurodevelopmental Channelopathies
- Tips, Tricks & Troubleshooting RNAscope® and BaseScope™ Assays
- Genetic Variant Detection in Cancer: Using RNAscope® ISH to Track Tumor Evolution
- CAR-T Cell Target Selection Using Innovative in Situ Hybridization Technology
- Getting Started and Troubleshooting Tips for RNAscope® and BaseScope™ Assays
- Combined immunohistochemistry and RNA in situ hybridization to elucidate tumor molecular heterogeneity in prostate cancer
- Characterization of immune checkpoint expression & infiltrating immune cells in TME by RNAscope ISH
- RNAscope® Assay Data Analysis Webinar
- Tips & Tricks to Get Publication Quality RNA ISH Data for Neuroscience Applications
- Product Introduction: RNAscope® VS Universal HRP, AP and Duplex Assays
- Get the most out of your RNAscope® Multiplex Fluorescent assays
- Optimize Your RNAscope® Assay Data Analysis
- Tips, Tricks & Troubleshooting to Get Publication Quality RNA ISH Data
- RNAscope® Assay Data Analysis Webinar
- BaseScope™ Assay: Recent Applications & FAQs
- Introduction to RNAscope® Probes
- New Product Introduction: RNAscope® Multiplex Fluorescent v2 Assay
- New Product Introduction: RNAscope® Multiplex Fluorescent v2 Assay
- Introduction to the RNAscope® Assay
- Visualize your research breakthroughs with RNAscope® and BaseScope™ assays
- Steps for Success with the RNAscope® Multiplex Fluorescent Assay
- RNAscope® Assay Data Interpretation
- Tips for Getting Started with the RNAscope® Multiplex Fluorescent Manual Assay
- Elucidating tumor heterogeneity in prostate cancer by combined IHC & RNA ISH
- New product introduction - BaseScope™ Assay, a new and innovative RNA ISH platform for the detection of short targets and exon junctions.
- New product introduction - BaseScope™ Assay, a new and innovative RNA ISH platform for the detection of short targets and exon junctions.
- New Product Introduction: RNAscope® LS Multiplex Fluorescent Assay
- Tips for Getting Started with the RNAscope® Multiplex Fluorescent Manual Assay
- Advanced Cell Diagnostics’ Pharma Assay Services – Expert capabilities in RNAscope ISH assays and analysis for detection and quantification of mRNA, transcript isoforms and point mutations.
- RNA-ISH for long non-coding RNAs (lncRNAs) - novel game changers in pathology
- Steps for Success with RNAscope® 2.5 VS on the Discovery Ultra and Discovery XT Automated Assays
- 2016 June 21st: Steps for Success with RNAscope® Assays & New Product Development
- Current State-of-the-Art Tissue-Based RNA Analysis with RNAscope Technology & Quantitative Analysis with the HALO Image Analysis Platform
- 2016 May17th: Become an RNAscope® Expert: RNAscope® Data Interpretation Webinar
- 2016 April 26: RNAscope® New Product Development - RNAscope® 2.5 HD Duplex Assay
- 2016 April 26: RNAscope® New Product Development - RNAscope® 2.5 HD Duplex Assay
- 2016 March 15th: New RNAscope® LS Duplex Assay
- 2016 Feb 26th: Become an RNAscope® Expert with the RNAscope® Multiplex Fluorescent assay
- 2016 Jan 28th: PTEN mRNA detection by chromogenic RNA in situ technologies: A reliable alternative to PTEN immunohistochemistry
- Steps for Success with the RNAscope® Assay & New Product Development
- Dec 17th, 2015: Steps for Success with RNAscope® Assay and New Product Development
- 2014 Sept 11th: Whole-Mount Zebrafish Embryos - RNAscope® for High-Resolution Detection of Transcripts with Localization of Proteins
- RNAscope® Technology: A Novel RNA in situ Hybridization Research Platform
- RNAscope® New Product Development - learn about RNAscope 2.5 product line
- RNAscope® New Product Development - learn about RNAscope 2.5 product line
- Linking Viral Discovery with Causality
- Dec 17th, 2015: Steps for Success with RNAscope® Assay and New Product Development
- Visualization of exogenous RNA using RNAscope ISH in rat brain to assess viral vector transduction
- Elucidating tumor heterogeneity in prostate cancer by combined IHC & novel RNA ISH
- Steps for success with the RNAscope assay: Basic training for new users
- Steps for success with the RNAscope assay: Basic training for new users
- Steps for success with the RNAscope assay: Basic training for new users
Follow a step-by-step visual guide to assist with running the RNAscope Manual Assay.
How to use EZ-Batch Tray
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 | |
How can we help you?
Please contact us directly, by phone or email, for any technical assistance with the RNAscope® assay or relevant products.
1 (877) 576-3636
