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. 2015:1189:123-32.
doi: 10.1007/978-1-4939-1164-6_9.

Active cell and ECM movements during development

Anastasiia Aleksandrova  1 , Brenda J Rongish, Charles D Little, András Czirók
Affiliations

Active cell and ECM movements during development

Anastasiia Aleksandrova et al. Methods Mol Biol. 2015.

Abstract

Dynamic imaging of the extracellular matrix (ECM) and cells can reveal how tissues are formed. Displacement differences between cells and the adjacent ECM scaffold can be used to establish active movements of mesenchymal cells. Cells can also generate large-scale tissue movements in which cell and ECM displacements are shared. We describe computational methods for analyzing multi-spectral time-lapse image sequences. The resulting data can distinguish between local "active" cellular motion versus large-scale, tissue movements, both of which occur during organogenesis. The movement data also provide the basis for construction of realistic biomechanical models and computer simulations of in vivo tissue formation.

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Figures

Fig. 1
Fig. 1
Image processing workflow to analyze cell and ECM displacements. Image pairs characteristic for a certain developmental stage are selected. The two sets of images were acquired in multiple focal (Z) planes and multiple microscopy modes. By a manual masking procedure, image regions are selected that contain either the region of interest or the anatomical reference used later. PIV analysis gives displacement estimation for the various optical modes, thus for the investigated cell population and ECM. The PIV data are weighted by the local standard deviation of the immunofluorescence intensity, which is used as a reliability score. Displacements obtained from a region of interest are registered to an anatomical frame of reference by subtracting displacement vectors characterizing an anatomical reference. Local tissue movements are established from ECM displacements by filtering techniques. Finally, the comparison of cell and local tissue movements yields cell-autonomous displacements
See this image and copyright information in PMC

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