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Related Experiment Video

Updated: Jan 11, 2026

High-throughput Imaging and Analysis Workflow for Evaluating Skin Cell Phenotypes and Proliferation States in Tissue Samples

High-throughput Imaging and Analysis Workflow for Evaluating Skin Cell Phenotypes and Proliferation States in Tissue Samples

Published on: October 31, 2025

High-throughput Imaging and Analysis Workflow for Evaluating Skin Cell Phenotypes and Proliferation States in Tissue

Salome Stierli¹, Flurin Sturzenegger², Whitney Shannon Jordaan³

¹Institute of Anatomy, University of Zurich; salome.stierli@anatomy.uzh.ch.

Journal of Visualized Experiments : Jove

|November 17, 2025

Summary

This study enhances skin regeneration research by combining the iterative-bleaching-extends-multiplexity (IBEX) method with Click-iT EdU labeling to accurately identify dividing cells in injured skin tissue.

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Last Updated: Jan 11, 2026

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Area of Science:

Regenerative medicine
Tissue engineering
Cell biology

Background:

Skin regeneration involves complex cellular interactions, with peripheral glial cells crucial for repair.
Current technological limitations hinder a complete understanding of these regenerative processes.
Accurate identification of proliferating cells is essential for studying tissue repair.

Purpose of the Study:

To characterize cellular composition changes during skin regeneration.
To overcome limitations in detecting dividing cells in skin tissue.
To develop an advanced imaging and analysis protocol for high-resolution tissue studies.

Main Methods:

Iterative-bleaching-extends-multiplexity (IBEX) method combined with Click-iT EdU chemistry.
High-throughput imaging using a fully automated spinning disk confocal microscope.
Development of an open-source image processing pipeline for data analysis.

Main Results:

The adapted IBEX method accurately identifies and categorizes actively dividing cell types in intact skin.
The combined approach overcomes underestimation of cell proliferation by traditional markers.
A novel, user-friendly image processing pipeline facilitates analysis of highly multiplexed imaging data.

Conclusions:

The IBEX protocol is versatile and adaptable to conventional imaging platforms for studying tissue regeneration.
Combining IBEX with Click-iT EdU labeling provides a powerful tool for dissecting dynamic cellular processes in skin.
This protocol advances the study of skin repair by enabling detailed spatial and temporal analysis of cell proliferation.