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Related Experiment Video

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Intravital Imaging of the Mouse Popliteal Lymph Node
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Diffusion of cytokines in live lymph node tissue using microfluidic integrated optical imaging.

A E Ross1, R R Pompano1

  • 1Department of Chemistry, University of Virginia, McCormick Rd., PO Box 400319, Charlottesville, VA 22904, USA.

Analytica Chimica Acta
|January 1, 2018
PubMed
Summary

We developed Microfluidic Integrated Optical Imaging (micro-IOI) to measure protein diffusion in live tissues. This novel method accurately quantifies cytokine transport, advancing our understanding of immune cell communication and immunotherapy design.

Keywords:
InflammationInterferon gammaInterleukin-2Local deliveryTumor necrosis factor alpha

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Area of Science:

  • Biophysics
  • Immunology
  • Microfluidics

Background:

  • Cellular communication and drug efficacy depend on protein diffusion in tissues.
  • Existing methods for analyzing tissue diffusion are invasive and can alter local transport properties.
  • Accurate measurement of protein diffusion in live tissue is crucial for understanding biological processes.

Purpose of the Study:

  • To develop and validate a novel, user-friendly method for quantifying bioactive protein diffusion in live tissue ex vivo.
  • To establish Microfluidic Integrated Optical Imaging (micro-IOI) as a reliable tool for studying transport phenomena in biological matrices.
  • To investigate the diffusion of various cytokines within live lymph node tissue.

Main Methods:

  • Developed a microfluidic platform for precise delivery of fluorescently labeled cytokines to microscale regions within live tissue slices.
  • Utilized widefield fluorescence microscopy to monitor protein diffusion.
  • Validated the micro-IOI method against theoretical predictions (Stokes-Einstein) and established techniques like fluorescence recovery after photobleaching (FRAP).

Main Results:

  • Micro-IOI demonstrated high accuracy, with free diffusion coefficients within 8% and 24% of theoretical predictions for dextrans and cytokines, respectively.
  • Diffusion coefficients measured by micro-IOI for dextrans and proteins in a model matrix were within 1.5-fold of FRAP results.
  • Successfully quantified the effective diffusion of three distinct cytokines (TNF-α, IFN-γ, IL-2) from different expression systems in live lymph node tissue.

Conclusions:

  • Microfluidic Integrated Optical Imaging (micro-IOI) is the first method to directly measure cytokine transport in live tissue slices.
  • This technique offers a non-invasive and user-friendly approach to study protein diffusion in complex biological environments.
  • The micro-IOI method has the potential to deepen the understanding of cell-cell communication dynamics and facilitate the design of targeted immunotherapies.