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Related Experiment Video

Updated: May 25, 2026

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Integrating Continuous Transepithelial Flux Measurements into an Ussing Chamber Set-Up.

Çlirim Alija1, Lukas Knobe1, Ioanna Pouyiourou1

  • 1Clinical Physiology/Nutritional Medicine, Medical Department, Division of Gastroenterology, Infectiology, Rheumatology, Charité-Universitätsmedizin Berlin, 12203 Berlin, Germany.

International Journal of Molecular Sciences
|February 24, 2024
PubMed
Summary

This study introduces a novel device for continuous monitoring of fluorescent compound flux across epithelial tissues. This method enables real-time assessment of paracellular permeability changes, overcoming limitations of traditional sampling techniques.

Keywords:
Ussing chamberautomatized flux measurementclaudinosmotic stressparacellular fluorescent markertight junctiontransepithelial flux

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Area of Science:

  • Epithelial physiology
  • Biophysical techniques
  • Pharmacology

Background:

  • Fluorescent compounds are crucial for studying epithelial paracellular transport.
  • Conventional flux measurements using sampling are inadequate for detecting rapid permeability shifts.
  • Existing methods lack real-time monitoring capabilities in Ussing chamber setups.

Purpose of the Study:

  • To develop and validate a device for continuous transepithelial flux measurements of fluorescent compounds.
  • To enable simultaneous recording of electrical and flux parameters in Ussing chambers.
  • To assess the device's utility in evaluating paracellular barrier function and permeability dynamics.

Main Methods:

  • Development of a flow-through chamber integrated with LED, optical filter, and photodiode for real-time fluorescence detection.
  • Calibration of the device using fluorescein across a concentration range of 1 nM to 150 nM.
  • Application of the device to various cell lines to measure fluorescein flux under different conditions, including altered barrier function and osmotic challenges.

Main Results:

  • The device demonstrated a linear output correlating with fluorescein concentration, indicating reliable calibration.
  • Fluorescein flux measurements were sensitive to changes in paracellular barrier integrity, modulated by tight junction protein expression and EGTA treatment.
  • Transcellular chloride secretion and osmotic manipulations significantly impacted transepithelial fluorescein flux, highlighting the device's ability to capture dynamic changes.

Conclusions:

  • The developed device provides a robust platform for continuous, real-time monitoring of fluorescent compound transepithelial flux.
  • This technology overcomes the limitations of traditional sampling methods, offering enhanced insights into epithelial barrier function.
  • The device's integration with Ussing chamber electrical recordings allows for comprehensive analysis of epithelial transport processes.