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Image analysis of lymphatic capillary junctions

MC Madeline J. Churchill
HB Haley du Bois
TH Taylor A. Heim
TM Tenny Mudianto
MS Maria M. Steele
JN Jeffrey C. Nolz
AL Amanda W. Lund
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Maximum projections of whole-mount z-stacks were converted into TIFF files using Zen Black (Zeiss). Images were blinded to the analyzer. The area of each VE-cadherin positive structure (cell junction) within single LYVE-1+ lymphatic capillaries was determined using the 3D Objects counter feature in Fiji (ImageJ; National Institutes of Health), irrespective of capillary area. The average size of junctions per capillary was calculated from each image and at least 10 capillaries averaged from each animal. Individual button junctions (punctate and not colocalized with LYVE-1) manually identified in naive capillaries have an average area of 0.53 ± 0.37 µm2, and zipper junctions are 3.5 ± 1.5 µm2 in area (Fig. S1, A and B).

Copyright and License information:  ©2022 Churchill et al.
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