• Oct
    23
    Single-Cell Transcriptomic Atlas of the Human Retina Identifies Cell Types Associated with Age-Related Macular Degeneration
    Madhvi Menon, PhD, Postdoctoral Research Fellow, Harvard Medical School and Brigham & Women’s Hospital
    Courtney Anderson, Applications Leader & Senior Scientist, Advanced Cell Diagnostics
    Single-Cell Transcriptomic Atlas of the Human Retina Identifies Cell Types Associated with Age-Related Macular Degeneration
    Madhvi Menon, PhD
    Postdoctoral Research Fellow, Harvard Medical School and Brigham & Women’s Hospital
    Courtney Anderson
    Applications Leader & Senior Scientist, Advanced Cell Diagnostics
    Outline:

    Age-related macular degeneration (AMD) is one of the leading causes of blindness in the elderly. Although genome-wide association studies have identified genetic variants associated with AMD, it has been challenging to identify the cell types driving this genetically complex disease. We performed single-cell RNA sequencing (scRNA-seq) of normal human retinas, and report the first single-cell human retinal transcriptomic atlas identifying all major cell types. Gene-expression heterogeneity was observed in Müller glial cells, suggesting that human retinal glia are more diverse than previously thought. The activity of genes associated with AMD was significantly enriched in vascular and glial cells, as well as in subpopulations of neurons. Furthermore, AMD disease-associated gene activity was consistent with regional macular disease susceptibility. Findings were validated using in situ hybridization techniques and immunofluorescence. Our data show how scRNA-seq can provide insight into relevant cell types for a complex genetic disease. 

    Learning Objectives:
    • Single-cell transcriptomic atlas of the human retina
    • Identification of cell types associated with AMD
    • Validation of findings using RNAscope®
    Dates and Registration:
    October
    23
    11:30am PST
    Register Now
  • Oct
    29
    RNAscope/BaseScope Probe Design Overview Webinar
    Outline:

    RNAscope/BaseScope Probe Design Overview Webinar

    Outline: Taking advantage of the RNAscope/BaseScope probe design strategies to achieve highly specific and sensitive detection of a variety of RNA targets in situ, including but not limited to RNA, mRNA, pre-RNA, TCR/BCR, KO validation, splice variant, circRNA, and fusion transcript.

    Who should attend:

    • Researchers interested in an overview of RNAscope/BaseScope probe design strategies.
    • Researchers interested in knowing the scope of targets that can be specifically detected by RNAscope/BaseScope probes.
    • Researchers interested in submitting standard probe design requests through ACD on-line portal.
    • Researchers interested in submitting non-standard probe design requests through ACD on-line portal.

    Learning Objectives:

    • Introduction to the RNAscope/BaseScope design workflow.
    • Featured applications of RNAscope/BaseScope probes including detection of mRNA, pre-RNA, KO event, TCR/BCR clone, splice variant, circRNA, gene fusion
    • Frequently asked questions - Applications (FAQs)
    • Step-by-step instructions and tips on filling the on-line new probe request (NPR) form.
    Dates and Registration:
    October
    29
    9:00 am PT
    12:00 PM ET
    6:00 pm CT
    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.

PresentationsDownload 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

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