Abstract
Organogenesis involves extensive and dynamic changes of tissue shape during development. It is associated with complex morphogenetic events that require enormous tissue plasticity and generate a large variety of transient three-dimensional geometries that are achieved by global tissue responses. Nevertheless, such global responses are driven by tight spatio-temporal regulation of the behaviours of individual cells composing these tissues. Therefore, the development of image analysis tools that allow for extraction of quantitative data concerning individual cell behaviours is central to study tissue morphogenesis. There are many image analysis tools available that permit extraction of cell parameters. Unfortunately, the majority are developed for tissues with relatively simple geometries such as flat epithelia. Problems arise when the tissue of interest assumes a more complex three-dimensional geometry. Here, we use the endothelium of the developing zebrafish dorsal aorta as an example of a tissue with cylindrical geometry and describe the image analysis routines developed to extract quantitative data on individual cells in such tissues, as well as the image acquisition and sample preparation methodology.This article is part of the Theo Murphy meeting issue 'Mechanics of development'.
Highlights
Tissue morphogenesis is inherent to embryonic development as tissues have to constantly and continuously adapt their shape during this process
Tooth bud formation or optic cup morphogenesis both involve very specific cell behaviours that have not been described in other developmental contexts [10,11]
To further advance our understanding of tissue morphogenesis, we must be able to follow individual cell behaviours in tissues with more complex 3D geometries. This has generated a demand for better 3D image analysis tools, which has been fuelled by the recent advancements in microscopy techniques in general and in live-imaging of whole tissues and organisms in particular [21]
Summary
Tissue morphogenesis is inherent to embryonic development as tissues have to constantly and continuously adapt their shape during this process. To further advance our understanding of tissue morphogenesis, we must be able to follow individual cell behaviours in tissues with more complex 3D geometries This has generated a demand for better 3D image analysis tools, which has been fuelled by the recent advancements in microscopy techniques in general and in live-imaging of whole tissues and organisms in particular [21]. Other approaches have focused on reducing data dimensionality in order to deal with the processing of large datasets produced by fast-acquisition imaging methods such as light-sheet microscopy [40,41] Such advances have enabled the study of tissue morphogenesis on organ or embryo surfaces that have arbitrary shapes as well as the disentanglement of signal from different layers [40].
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