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Computational and Mathematical Methods in Medicine
Volume 2014, Article ID 820214, 9 pages
http://dx.doi.org/10.1155/2014/820214
Research Article

Automated Image Analysis of Lung Branching Morphogenesis from Microscopic Images of Fetal Rat Explants

1ICVS/3B’s, PT Government Associate Laboratory, Guimarães, Braga, Portugal
2DIGARC, Polytechnic Institute of Cávado and Ave, Barcelos, Portugal
3Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, 4710-057 Braga, Portugal

Received 28 May 2014; Accepted 21 July 2014; Published 28 August 2014

Academic Editor: Carlo Cattani

Copyright © 2014 Pedro L. Rodrigues et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background. Regulating mechanisms of branching morphogenesis of fetal lung rat explants have been an essential tool for molecular research. This work presents a new methodology to accurately quantify the epithelial, outer contour, and peripheral airway buds of lung explants during cellular development from microscopic images. Methods. The outer contour was defined using an adaptive and multiscale threshold algorithm whose level was automatically calculated based on an entropy maximization criterion. The inner lung epithelium was defined by a clustering procedure that groups small image regions according to the minimum description length principle and local statistical properties. Finally, the number of peripheral buds was counted as the skeleton branched ends from a skeletonized image of the lung inner epithelia. Results. The time for lung branching morphometric analysis was reduced in 98% in contrast to the manual method. Best results were obtained in the first two days of cellular development, with lesser standard deviations. Nonsignificant differences were found between the automatic and manual results in all culture days. Conclusions. The proposed method introduces a series of advantages related to its intuitive use and accuracy, making the technique suitable to images with different lighting characteristics and allowing a reliable comparison between different researchers.