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Related Concept Videos

Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry
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Multiplex Immunohistochemistry and Immunofluorescence: A Practical Update for Pathologists.

Paul W Harms1, Timothy L Frankel2, Myrto Moutafi3

  • 1Department of Pathology, Michigan Medicine/University of Michigan, Ann Arbor, Michigan; Department of Dermatology, Michigan Medicine/University of Michigan, Ann Arbor, Michigan; Rogel Cancer Center, Michigan Medicine/University of Michigan, Ann Arbor, Michigan.

Modern Pathology : an Official Journal of the United States and Canadian Academy of Pathology, Inc
|April 27, 2023
PubMed
Summary
This summary is machine-generated.

Multiplex staining techniques offer advanced tissue analysis for improved cancer diagnosis and immuno-oncology research. These methods enable simultaneous evaluation of multiple biomarkers, overcoming limitations of traditional single-marker approaches in clinical practice.

Keywords:
digital pathologyimmunotherapymultiplex immunofluorescencemultiplex immunohistochemistrytumor microenvironment

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Area of Science:

  • Oncology
  • Immunology
  • Pathology
  • Digital Health

Background:

  • Advances in understanding the tumor microenvironment and antitumor immunity have driven progress in cancer immunotherapy.
  • Current tissue-based diagnostics rely on traditional methods like hematoxylin and eosin staining and single-biomarker immunohistochemistry (IHC), which are insufficient for complex analyses.
  • Digital pathology offers new tools for tumor microenvironment assessment, but traditional methods limit simultaneous evaluation of multiple biomarkers.

Purpose of the Study:

  • To review different multiplex staining techniques for immunohistochemistry (IHC) and immunofluorescence.
  • To discuss alternatives to multiplex staining, such as epitope-based tissue mass spectrometry and digital spatial profiling (DSP).
  • To highlight the potential of multiplex staining for improving diagnostic practice and immuno-oncology research.

Main Methods:

  • Description of various multiplexed IHC and immunofluorescence approaches.
  • Discussion of alternative techniques including epitope-based tissue mass spectrometry, digital spatial profiling (DSP), and virtual multiplexing.
  • Emphasis on the need for standardized protocols and digital pathology workflows for analyzing multiplexed stains.

Main Results:

  • Multiplex staining techniques are technically feasible for most clinical laboratories.
  • Traditional single-biomarker assessments are poorly suited for complex quantitative analyses.
  • Alternative methods like DSP require specialized platforms, limiting accessibility.

Conclusions:

  • Multiplex staining holds significant potential to enhance diagnostic practices and immuno-oncology research.
  • Standardization and digital pathology workflows are crucial for effective analysis of multiplexed stains.
  • Multiplex staining is poised for routine clinical diagnostic use in the near future.