An integrated pipeline for the multidimensional analysis of branching morphogenesis

Combes, Alexander N., Short, Kieran M., Lefevre, James, Hamilton, Nicholas A., Little, Melissa H. and Smyth, Ian M. (2014) An integrated pipeline for the multidimensional analysis of branching morphogenesis. Nature Protocols, 9 12: 2859-2879. doi:10.1038/nprot.2014.193


Author Combes, Alexander N.
Short, Kieran M.
Lefevre, James
Hamilton, Nicholas A.
Little, Melissa H.
Smyth, Ian M.
Title An integrated pipeline for the multidimensional analysis of branching morphogenesis
Journal name Nature Protocols   Check publisher's open access policy
ISSN 1750-2799
1754-2189
Publication date 2014-11-20
Sub-type Article (original research)
DOI 10.1038/nprot.2014.193
Open Access Status
Volume 9
Issue 12
Start page 2859
End page 2879
Total pages 21
Place of publication London, United Kingdom
Publisher Nature Publishing Group
Collection year 2015
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-2â €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 across developmental time. The protocol establishes a benchmark for assessing the impact of genetic change or fetal environment on organogenesis that does not rely on ex vivo organ culture or section-based reconstruction. By using this approach, examination of two developmental stages for an organ such as the kidney can be undertaken by a postdoctoral-level researcher in 6 weeks, with a full developmental analysis in mouse achievable in 5 months.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
Institute for Molecular Bioscience - Publications
 
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