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Related Concept Videos

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research09:12

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research

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This report describes an automated protocol for multiplex immunofluorescence (mIF) assays on an automated slide stainer. It demonstrates the high reproducibility of spatial proteomics on standard high-throughput tissue autostainers and whole-slide fluorescence imagers, which is ideal for custom mIF assays across a large number of tissue slides in translational...
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Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment06:05

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In this article, a protocol for manual tyramide signal amplification (TSA) multiplex immunofluorescence (mIF) combined with image analysis and spatial analysis is described. This protocol can be used with formalin-fixed paraffin-embedded (FFPE) sections for the staining of two to six antigens per slide depending on the slide scanner available in the...
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A detailed protocol for a six-marker multiplex immunofluorescence panel is optimized and performed, using an automated stainer for more consistent results and a shorter procedure time. This approach can be directly adapted by any laboratory for immuno-oncology...
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Multiplex assays can provide beneficial information for basic cellular mechanisms and eliminate waste of reagents and unnecessary repetitive experiments. We describe here a multiplex caspase-3/7 activity assay, using fluorescent- and luminescent-based methods, to determine cell viability in an in vitro hypothalamic model following oxidative challenge with palmitic...
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Related Experiment Video

Updated: Jan 19, 2026

High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research
09:12

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Published on: June 10, 2025

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Multiplex Immunofluorescence Assays.

Alejandro Francisco-Cruz1,2, Edwin Roger Parra1,2, Michael T Tetzlaff1,2

  • 1Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 11, 2019
PubMed
Summary

Multiplexed imaging enhances the study of tumor immune contexture using formalin-fixed, paraffin-embedded tissues. This technology aids in understanding the tumor microenvironment and identifying biomarkers for treatment and translational research.

Keywords:
Cancer tissueDigital image analysisImmune-profilingTyramide signal amplification

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Area of Science:

  • Biomedical Imaging
  • Oncology
  • Immunology

Background:

  • Multiplexed imaging platforms enable simultaneous detection of multiple epitopes on single tissue sections.
  • These technologies are crucial for studying tumor immune contexture and the tumor microenvironment.
  • They offer advanced methods for evaluating formalin-fixed and paraffin-embedded (FFPE) tissues.

Purpose of the Study:

  • To review multiplexed imaging techniques for FFPE tumor tissues.
  • To focus on multiplex immunofluorescence with tyramine signal amplification for immune profiling.
  • To highlight applications in translational research and biomarker discovery.

Main Methods:

  • Utilizing multiplexed imaging platforms for simultaneous antigen detection.
  • Applying digital image analysis software for high-throughput assays.
  • Employing multiplex immunofluorescence with tyramine signal amplification staining.

Main Results:

  • Improved characterization of cell populations and their spatial distribution in tissues.
  • Enhanced understanding of tumor microenvironment and its relation to clinical outcomes.
  • Facilitation of prognostic and predictive biomarker identification.

Conclusions:

  • Multiplexed imaging is a powerful tool for in-depth tumor evaluation.
  • It significantly advances immune profiling and translational research in oncology.
  • These technologies are revolutionizing the study of cancer specimens.