• Jun
    05
    2025 June 5 - Multiomics Image Analysis-Driven Insights into Tumor Microenvironment
    Vytautas Navikas, PhD, , Data Scientist, Lunaphore Technologies
    2025 June 5 - Multiomics Image Analysis-Driven Insights into Tumor Microenvironment
    Vytautas Navikas, PhD,
    Data Scientist, Lunaphore Technologies
    Outline:

    Glioblastoma (GBM), known as one of the most aggressive primary neuroepithelial tumors, presents significant challenges in diagnosis and treatment. Traditional immunohistochemistry often falls short in capturing the intricate tumour microenvironment (TME) complexity within GBM. With an increasing emphasis on biomarker discovery and targeting complex diseases like GBM, spatial biology has emerged as a fundamental approach, revealing multiple biomarkers while preserving spatial tissue information. The pivotal task of transforming information-rich multiplex images into crucial quantitative data remains a significant hurdle. In this webinar, we present Lunaphore's cutting-edge solutions for generating and analyzing hyperplex images. 

    The portfolio encompasses COMET™, a fully-automated spatial multiomics platform, and HORIZON™, an image analysis software tailored for COMET™ hyperplex immunofluorescence images. COMET™ integrates RNAscope™ HiPlex Pro technology with a sequential immunofluorescence (seqIF™) approach to simultaneously detect RNA and proteins on a single tissue section. Here, a multiomics panel featuring 12 RNAs and 24 proteins reveals insights into immune and cancer cell populations and their cellular activity within a TME.  

    To show the full potential of multiomics data analytics, the acquired datasets were analyzed using HORIZON™. It enables the performance of a single-cell analysis workflow for deep-cell phenotyping while using multiomics data to get multi-dimensional insights about the sample. The batch analysis was performed on multiple regions of interest to apply the same analysis workflow, encompassing AI-driven cell segmentation, followed by RNA signal detection, feature extraction, and supervised classification. Additionally, spatial evaluations of cellular phenotypes of interest unveiled intricate organizational relationships within the TME and provided valuable hypotheses-driven insights into the sample.

    This webinar showcases how spatial multiomics offers a comprehensive view of the TME, advancing  our comprehension of GBM pathology. Leveraging the single-cell resolution of spatial multiomics on COMET™ and intuitive image analysis with HORIZON™, researchers can seamlessly navigate the complex immune and cancer networks within the TME.

    Dates and Registration:
    June
    05
    3pm CEST
    Register Now

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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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