RNAscope Webinars, Videos, Presentations | In Situ Hybridization, RNA-ISH

Jun
25
Spatial Mapping of Gene Expression at Single Cell Resolution with the RNAscope^® Technology
Courtney Anderson, Ph.D., , Research Manager, Applications
Spatial Mapping of Gene Expression at Single Cell Resolution with the RNAscope^® Technology
Courtney Anderson, Ph.D.,
Research Manager, Applications
Outline:
Complex, highly heterogeneous tissues, such as the brain, tumors, and developing organs, are comprised of multiple cell types and states. However, interrogation of these complex tissue types requires a highly sensitive, specific, and multiplexed spatial approach with single cell resolution.
Learning Objectives:
In this webinar we will show how researchers can use the RNAscope^® in situ hybridization technology to:
- Visualize the cellular heterogeneity of complex organs with the multiplexing and spatial capabilities of the RNAscope^® Multiplex Fluorescent assay
- Spatially map scRNA-seq gene profiles at the single cell level in the tissue context
- Localize newly identified cell subtypes and cell markers
- Confirm scRNA-seq results from publicly available datasets in the tissue context
Dates and Registration:
June
25
9:00 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.

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