Extended Data Fig. 3: Imaging-based assessment and quantification of the post-natal cardiac lymphatic plexus.
From: Cardiac lymphatics retain LYVE-1-dependent macrophages during neonatal mouse heart regeneration
Light-sheet and wholemount confocal imaging of Prox1-tdTomato (A-C) and Vegfr3+/lacZ (D) intact hearts, at post-natal days 7 and 14 respectively, enable visualisation of the cardiac lymphatic plexus which can be segmented using Image J (E) and quantified in terms of parameters such as vessel length, calibre, junctions, branch points etc using AngioToolTM (F). The total vessel length and number of end points remained stable until P14 and then significantly increased on both sides of the heart via lymphangiogenesis (G). By P16 the lymphatics were fully formed on the dorsal side, while the ventral lymphatics were delayed, reaching full expansion by P21 (H). Data are presented as mean ± SEM. n = 5 for P1, P16, P21; n = 3 for P3, P5, P9; n = 4 for P8, P11, P28. Significant differences were calculated using 2-way ANOVA. Scale bars 0.5 mm, A and 1 mm, D.
