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Related Experiment Video

Updated: Sep 18, 2025

Non-invasive Optical Imaging of the Lymphatic Vasculature of a Mouse
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Semi-Automated Method for Analyzing Lymphatic Vessels From Immunofluorescence Images.

Victor Rojas-Henríquez1, Alexandra Olate-Briones1, Celia Salazar1

  • 1Lymphatic Vasculature and Inflammation Research Laboratory, Facultad de Ciencias de la Salud, Instituto de Ciencias Biomédicas, Universidad Autónoma de Chile, Talca, Chile.

Current Protocols
|June 25, 2025
PubMed
Summary

A new semi-automated protocol accurately quantifies lymphatic vasculature (LV) coverage in tissues. This method improves efficiency and precision for analyzing LV changes during inflammation, crucial for understanding immune cell trafficking.

Keywords:
colonlymphatic vasculaturemeasurement areameningeal lymphatic vasculature

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Area of Science:

  • Immunology
  • Cell Biology
  • Microscopy

Background:

  • Lymphatic vasculature (LV) is vital for immune cell trafficking and its dysfunction is linked to inflammatory diseases.
  • Accurate assessment of LV coverage is essential for quantifying inflammation.
  • Manual measurement of LV coverage is time-consuming and prone to errors.

Purpose of the Study:

  • To develop an improved, semi-automated protocol for determining lymphatic vasculature coverage area in intestinal and meningeal tissues.
  • To enhance the accuracy and efficiency of LV analysis compared to manual methods.

Main Methods:

  • Developed a semi-automated protocol using Fiji software's Threshold tool for LV analysis.
  • Applied the protocol to mouse colon tissue and meninges stained with LV-specific antibodies.
  • Compared the semi-automated protocol with manual selection methods.

Main Results:

  • The semi-automated protocol significantly improved accuracy and efficiency in measuring LV coverage.
  • Automated selection minimized operator errors and time investment.
  • Achieved more specific selections and accurate measurements than manual methods.

Conclusions:

  • The developed semi-automated protocol offers a more accurate and efficient method for assessing lymphatic vasculature coverage.
  • This protocol aids in the precise analysis of LV changes during inflammation.
  • The protocol is adaptable to other microscopy techniques with sufficient image contrast and known scale.