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Related Experiment Video

Updated: May 2, 2026

High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence
10:28

High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence

Published on: October 28, 2025

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High-Plex Tissue Imaging with Conventional Immunofluorescence Platforms and Open-Source Software via Iterative

Ian T Lamborn1, Hiroshi Ichise1,2, Emily Speranza3

  • 1Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|April 30, 2026
PubMed
Summary
This summary is machine-generated.

Iterative bleaching extends multiplexity (IBEX) is a simple immunofluorescence technique for multiplex tissue imaging. This method is accessible for labs with limited immunohistochemistry experience.

Keywords:
IBEXImmunofluorescenceMicroscopyMultiplexOptical imagingSpatial proteomics

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Area of Science:

  • Biomedical imaging
  • Immunohistochemistry
  • Molecular biology

Background:

  • Multiplex immunofluorescence enables simultaneous detection of multiple targets in tissue samples.
  • High-plex imaging traditionally requires specialized equipment and expertise.
  • There is a need for accessible, user-friendly multiplex imaging methods.

Purpose of the Study:

  • To describe the implementation of Iterative Bleaching Extends Multiplexity (IBEX).
  • To provide a guide for laboratories with minimal immunohistochemistry experience.
  • To demonstrate the accessibility of advanced multiplex imaging techniques.

Main Methods:

  • Iterative Bleaching Extends Multiplexity (IBEX) protocol.
  • Utilizes standard microscopy platforms and commercial reagents.
  • Employs open-source software for image analysis.

Main Results:

  • IBEX is an easy-to-use method for highly multiplex immunofluorescent tissue imaging.
  • The protocol is adaptable for laboratories with limited prior experience.
  • Successful implementation is demonstrated with readily available resources.

Conclusions:

  • IBEX offers a practical solution for high-plex tissue imaging.
  • The method lowers the barrier to entry for multiplex immunofluorescence.
  • Facilitates broader adoption of advanced imaging techniques in research settings.