MENU
NOV 12, 2020 9:00 AM PST

Fluorescent multiplexing of 3D spheroid FFPE with Roche automated stainer and reagents: assay optimization, imaging and future implications

Speaker
  • Staff Scientist
    Biography

      Srabani Bhaumik is a Staff Scientist in the Strategic Applications and Innovation Lab at Roche Tissue Diagnostics. Her R&D experiences includes molecular imaging, molecular pathology, small animal imaging, MRI contrast media imaging, reporter gene imaging, regenerative medicine and in-vitro assay development in oncology. Her current work is focused on fluorescent multiplexing (mIF) assay development on Roche automated IHC/ISH instruments, image acquisition and image analysis on multiple platforms. She was the technical lead for the development and launch of the fluorescent RUO 5-Plex assay on Roche Discovery platform. She has successfully developed complex fluorescent multiplex assays on various oncology samples including 3D cell spheroids and cytology. Prior to Roche, she worked at General Electric Global Research, where she focused on the MultiOmyx platform to explore immune oncology biomarker.


    Abstract

    Three-dimensional (3D) cell culture spheroids and aggregates are preferred over monolayer cell culture due to their architectural and functional similarity to solid tumors. To study expression profiles of key tumor biomarkers, physiologically relevant preclinical models of non-small cell lung adenocarcinoma, and colon and colorectal adenocarcinoma cell line‐derived 3D spheroids and aggregates were utilized. We performed multiplex immunofluorescence (mIF) staining, whole slide scanning and image analysis to enable simultaneous detection and quantification of multiple biomarkers on a single formalin-fixed paraffin embedded section.
    We describe the workflow optimized in our lab at Roche Tissue Diagnostics to establish mIF assay for 3D spheroids and aggregates sections. Multiplex staining of 3D sections were performed on Roche DISCOVERY Ultra automated staining instruments with Roche reagent and fluorescent dyes. Biomarker panels were designed to examine the expression of frequently studies key markers of two cancer subtypes. The slides were scanned and multispectral images captured in Vectra 3 (Akoya Biosciences) fitted with filters compatible to the dyes. Quantitative assessment of the expressed biomarkers was performed through image analysis.


    Show Resources
    You May Also Like
    MAY 11, 2021 10:00 AM PDT
    Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
    C.E. CREDITS
    MAY 11, 2021 10:00 AM PDT
    Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
    Date: May 11, 2021 Time: 10:00zm PDT Your samples are some of the most valuable assets in the laboratory. After spending countless hours on extraction and preparation, your conclusions could...
    NOV 18, 2020 8:00 AM PST
    C.E. CREDITS
    NOV 18, 2020 8:00 AM PST
    DATE: November 18, 2020 TIME: 08:00am PDT We develop and implement technologies to solve some of the major bottlenecks in biomedical research. In particular, we establish new imaging approac...
    NOV 16, 2020 8:00 AM PST
    C.E. CREDITS
    NOV 16, 2020 8:00 AM PST
    Date: November 16, 2020 Time: 8:00am (PST), 11:00am (EST) CRISPR screening has become the prime discovery tool in modern biomedical research and drug discovery. At the same time, most screen...
    FEB 24, 2021 10:00 AM PST
    C.E. CREDITS
    FEB 24, 2021 10:00 AM PST
    DATE: February 24, 2021 TIME: 10am PST Automated lab instruments such as liquid handlers and cell sorters are increasingly common in all types of laboratories, driving fast results for labor...
    APR 01, 2021 8:00 AM PDT
    C.E. CREDITS
    APR 01, 2021 8:00 AM PDT
    Date: April 01, 2021 Time: 8:00am (PST), 11:00am (EST) Generating therapeutic antibodies is far more challenging than obtaining antibodies that merely recognize their targets. Engineering po...
    DEC 03, 2020 4:30 PM PST
    C.E. CREDITS
    DEC 03, 2020 4:30 PM PST
    This drug development program is designed to create a family of broad-spectrum, pan-coronaviral drugs that respectively inhibit multiple key enzymes required for viral replication. By target...
    Loading Comments...
    Show Resources