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  • May
    18
    2021 May 18: Use of RNAscope technology to study reproductive health and dysfunction in the rodent central nervous system
    Lique Coolen, Ph.D.,, Professor, Department Biological Sciences, Associate Dean College Arts and Sciences, Kent State University
    Aleisha Moore, Ph.D., Assistant Professor, Department Biological Sciences, Kent State University
    2021 May 18: Use of RNAscope technology to study reproductive health and dysfunction in the rodent central nervous system
    Lique Coolen, Ph.D.,
    Professor, Department Biological Sciences, Associate Dean College Arts and Sciences, Kent State University
    Aleisha Moore, Ph.D.
    Assistant Professor, Department Biological Sciences, Kent State University
    Outline:

    In this webinar, you will hear from Dr. Lique Coolen and Dr. Aleisha Moore from Kent State University on how they have integrated powerful RNAscope in situ hybridization technology in their research to study the mechanisms underlying spinal cord injury and neuroendocrine dysfunction.



    Dr. Lique Coolen will provide an overview of studies in her laboratory that examine the impact of spinal cord injury on the spinal reflex generator for sexual function in the lumbar spinal cord in male rats. Recent evidence of injury-induced changes in transcript levels of neuropeptides and identification of localization of receptors in this spinal generator using RNAscope in situ hybridization will be presented. Furthermore, pharmacological experiments towards treatment development will be introduced.



    Dr. Aleisha Moore will present work in her laboratory identifying changes in neuronal circuits controlling fertility that may lead to polycystic ovarian syndrome, the most common cause of infertility in women worldwide. In her talk, Dr. Moore will focus on how she used RNAscope HiPlex technology in a mouse model of PCOS to evaluate changes in gene expression patterns within complicated and heterogenous hypothalamic neuronal circuits that may underlie the pathogenesis of PCOS.

    Learning Objectives:

    Key learning objectives in Dr. Coolens talk:

    • Learn about impacts of spinal cord injury on urogenital function.
    • Become familiar with the use of RNAscope to identify G-protein coupled receptors in spinal cord.

     

    Key learning objectives in Dr. Moore’s talk:

    • Learn about the neuroendocrine pathogenesis of PCOS.
    • Learn about the current use of RNAscope HiPlex technology to study hypothalamic cell populations.
    • Become familiar with Cellprofiler software for automated analysis of RNAscope HiPlex data.
    Dates and Registration:
    May
    18
    8am PST
    11am EST
    5pm CEST
    Register Now
  • May
    20
    2021 May 20 - Using Spatial Transcriptomics to Understand the Molecular Mechanisms of Memory
    Kaitlin Sullivan, MSc, PhD Student, University of British Columbia, Cembrowski Lab
    Emily Martersteck, Field Application Scientist, Advanced Cell Diagnostics, a Bio-Techne Brand
    2021 May 20 - Using Spatial Transcriptomics to Understand the Molecular Mechanisms of Memory
    Kaitlin Sullivan, MSc
    PhD Student, University of British Columbia, Cembrowski Lab
    Emily Martersteck
    Field Application Scientist, Advanced Cell Diagnostics, a Bio-Techne Brand
    Outline:

    This webinar will present a research project that is using spatial transcriptomics to discern the molecular mechanisms underlying fear memory. 

    Kaitlin Sullivan of the University of British Columbia will discuss how she is using multiplexed, fluorescent RNA in situ hybridization (mFISH) based on the RNAscope technology to assess neuronal activity during the creation and recollection of fear memory.

    Learning Objectives:
    • The use of mFISH to spatially resolve transcriptomically unique neuronal subpopulations.
    • How to computationally analyze images and quantify expression at single-cell, single-molecule resolution.
    • How to pair mFISH with morphological, circuit, and functional information to get a holistic view of cell types across multiple spatial scales.
    Dates and Registration:
    May
    20
    9am PST
    12pm EST
    6pm CEST
    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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