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Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
07:52

Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma

Published on: January 9, 2019

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Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma

Guo Hong1, Shuangyi Fan2, The Phyu2

  • 1Department of Laboratory Medicine, AnSteel Group General Hospital; Cancer Science Institute of Singapore, National University of Singapore.

Journal of Visualized Experiments : Jove
|January 29, 2019
PubMed

Insights

This study presents a multiplexed fluorescent immunohistochemistry (IHC) protocol for lymphoma research. The method allows for quantitative assessment of multiple protein targets within specific cell types, improving diagnostic accuracy.

Area of Science:

  • Biomedical Sciences
  • Molecular Biology
  • Immunohistochemistry

Background:

  • Conventional chromogenic immunohistochemistry (IHC) faces challenges in visualizing and quantifying multiple antigens in a single tissue section.
  • Multiplexed imaging is crucial for lymphoma research and diagnostics due to the complex tumor microenvironment and the need to interpret multiple markers.

Purpose of the Study:

  • To describe a protocol for multiplexed fluorescent IHC staining for lymphoma research.
  • To enable quantitative assessment of multiple protein targets in specific cell types within lymphoma tissue.
  • To provide a method for improved spatial and quantitative analysis of the lymphoma microenvironment.

Main Methods:

  • Development and optimization of a multiplexed fluorescent IHC protocol.
  • Antibody validation and optimization for multiplex staining.
  • Staining of tissue microarray (TMA) slides with lymphoma markers.
  • Slide scanning and quantitative data analysis, including mean intensity and percentage positivity scores.

Main Results:

  • Successful implementation of a multiplexed fluorescent IHC protocol for lymphoma.
  • Generation of quantitative scores for marker intensity and percentage positivity.
  • Demonstration of minimized sample utilization and provision of spatial information for multiple markers.

Conclusions:

  • Multiplexed fluorescent IHC offers a powerful approach for quantitative analysis of multiple protein targets in lymphoma.
  • This method enhances the understanding of the lymphoma tumor microenvironment and aids in research and diagnostics.
  • The protocol facilitates accurate assessment of protein expression in specific cell types, leading to improved diagnostic capabilities.

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