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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
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.
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.
