TubuleTracker: a high-fidelity shareware software to quantify angiogenesis architecture and maturity
Journal:
arXiv
Published Date:
Jul 2, 2025
Abstract
Background: In vitro endothelial cell culture is widely used to study
angiogenesis. Histomicrographic images of cell networks are often analyzed
manually, a process that is time-consuming and subjective. Automated tools like
ImageJ (NIH) can assist, but are often slow and inaccurate. Additionally, as
endothelial networks grow more complex, traditional architectural metrics may
not fully reflect network maturity. To address these limitations, we developed
tubuleTracker, a software tool that quantifies endothelial network architecture
and maturity rapidly and objectively. Methods: Human umbilical vein endothelial
cells were cultured in an extracellular matrix, and 54 images were acquired
using phase contrast microscopy. Each image was analyzed manually by three
independent reviewers, and by both ImageJ and tubuleTracker. Key metrics
included tubule count, total length, node count, tubule area, and vessel
circularity. In parallel, trained scientists rated each image for angiogenesis
maturity on a 1-5 scale (1 = most mature). Results: Analysis time per image
differed significantly: manual (8 min), ImageJ (58+/-4 s), and tubuleTracker
(6+/-2 s) (p<0.0001). Significant differences were also found in tubule count
(manual 168+/-SD, tubuleTracker 92+/-SD, ImageJ 433+/-SD), length, and node
count (all p<0.0001). tubuleTracker's metrics varied significantly across
angiogenesis maturity scores, including tubule count, length, node count, area,
and circularity (all p<0.0001). Conclusions: tubuleTracker was faster and more
consistent than both manual and ImageJ-based analysis. Vessel circularity
proved especially effective in capturing angiogenesis maturity. tubuleTracker
is available as free shareware for the biomedical research community.
