Abstract
The lymphatic vasculature is critical for immunity and interstitial fluid homeostasis, playing important roles in diseases such as lymphedema and metastatic cancer. Animal models have been generated to explore the role of lymphatics and lymphangiogenic growth factors in such diseases, and to study lymphatic development. However, analysis of lymphatic vessels has primary been restricted to counting lymphatics in two-dimensional tissue slices, due to a lack of more sophisticated methodologies. In order to accurately examine lymphatic dysfunction in these models, and analyse the effects of lymphangiogenic growth factors on the lymphatic vasculature, it is essential to quantify the morphology and patterning of the distinct lymphatic vessels types in three-dimensional tissues. Here, we describe a method for performing such analyses, integrating user-operated image-analysis software with an approach that considers important morphological, anatomical and patterning features of the distinct lymphatic vessel subtypes. This efficient, reproducible technique is validated by analysing healthy and pathological tissues.
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Notes on contributors
Ramin Shayan
Author Contributions R. Shayan and T. Karnezis concept and method development, experimental work and manuscript preparation; E. Tsantikos and S.P. Williams experimental work and manuscript preparation; A.S. Runting software design and testing; M.W. Ashton, M.G. Achen, M.L. Hibbs, and S.A. Stacker concept and manuscript preparation.
Tara Karnezis
Author Contributions R. Shayan and T. Karnezis concept and method development, experimental work and manuscript preparation; E. Tsantikos and S.P. Williams experimental work and manuscript preparation; A.S. Runting software design and testing; M.W. Ashton, M.G. Achen, M.L. Hibbs, and S.A. Stacker concept and manuscript preparation.
Marc G. Achen
Competing Interest Statement S.A. Stacker and M.G. Achen are consultants for Vegenics Ltd.
Steven A. Stacker
Competing Interest Statement S.A. Stacker and M.G. Achen are consultants for Vegenics Ltd.