Journal article

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

Abstract

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..

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Grants

Awarded by National Health and Medical Research Council of Australia


Awarded by Australian Research Council


Awarded by Human Frontiers in Science Program


Funding Acknowledgements

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.