An integrated pipeline for the multidimensional analysis of branching morphogenesis
Alexander N Combes, Kieran M Short, James Lefevre, Nicholas A Hamilton, Melissa H Little, Ian M Smyth
NATURE PROTOCOLS | NATURE PUBLISHING GROUP | Published : 2014
Developmental branching morphogenesis establishes organ architecture, and it is driven by iterative interactions between epithelial and mesenchymal progenitor cell populations. We describe an approach for analyzing this interaction and how it contributes to organ development. After initial in vivo cell labeling with the nucleoside analog 5-ethynyl-2'-deoxyuridine (EdU) and tissue-specific antibodies, optical projection tomography (OPT) and confocal microscopy are used to image the developing organ. These imaging data then inform a second analysis phase that quantifies (using Imaris and Tree Surveyor software), models and integrates these events at a cell and tissue level in 3D space and acro..View full abstract
Awarded by National Health and Medical Research Council of Australia
Awarded by Australian Research Council
Awarded by Human Frontiers in Science Program
This work was supported by the National Health and Medical Research Council of Australia (APP1002748, APP1022239 and APP1063989), the Australian Research Council (DP10310086) and the Human Frontiers in Science Program (RGP0039/2011). Microscopy was performed at the Australian Cancer Research Foundation (ARCF)/Institute for Molecular Bioscience Cancer Biology Imaging Facility, which was established with the support of the ARCF. I.M.S. holds a Future Fellowship from the Australian Research Council. M.H.L. is a Senior Principal Research Fellow of the Australian National Health and Medical Research Council.