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Endothelial cells line the interior lumen of blood and lymphatic vessels. Tube formation assays recreate the formation of these vessels in vitro. This process is stimulated by the application of vascular endothelial growth factor (VEGF), which binds to the receptor VEGFR2. This protocol outlines the setup and time-lapse imaging of a tube formation assay using Livecyte, a live-cell, label-free, quantitative phase imaging (QPI) microscope. Subsequent software analysis workflows are then described to quantify the cells’ mass, motility, morphology, and network formation. Using a skeletonization process, the Livecyte software can quantify the number of nodes, segments, branches, junctions, mean segment length, and total network length over time, allowing researchers to determine the dynamic changes in network formation throughout the assay.

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Label-Free Imaging with Automated Tracking and Cell-by-Cell Analysis of Migration, Morphology, and Differentiation of Endothelial Cells

  • 实验方案

  • 发表日期：

    2025年

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  • Jessica Rickman,
  • Meetal Jotangia,
  • James Russell,
  • Martin Humphry

摘要

Endothelial cells line the interior lumen of blood and lymphatic vessels. Tube formation assays recreate the formation of these vessels in vitro. This process is stimulated by the application of vascular endothelial growth factor (VEGF), which binds to the receptor VEGFR2. This protocol outlines the setup and time-lapse imaging of a tube formation assay using Livecyte, a live-cell, label-free, quantitative phase imaging (QPI) microscope. Subsequent software analysis workflows are then described to quantify the cells’ mass, motility, morphology, and network formation. Using a skeletonization process, the Livecyte software can quantify the number of nodes, segments, branches, junctions, mean segment length, and total network length over time, allowing researchers to determine the dynamic changes in network formation throughout the assay.