Abstract
The need for quantification of cell growth patterns in a multilayer, multi-cellular tissue necessitates the development of a 3D reconstruction technique that can estimate 3D shapes and sizes of individual cells from Confocal Microscopy (CLSM) image slices. However, the current methods of 3D reconstruction using CLSM imaging require large number of image slices per cell. But, in case of Live Cell Imaging of an actively developing tissue, large depth resolution is not feasible in order to avoid damage to cells from prolonged exposure to laser radiation. In the present work, we have proposed an anisotropic Voronoi tessellation based 3D reconstruction framework for a tightly packed multilayer tissue with extreme z-sparsity (2–4 slices/cell) and wide range of cell shapes and sizes. The proposed method, named as the ‘Adaptive Quadratic Voronoi Tessellation’ (AQVT), is capable of handling both the sparsity problem and the non-uniformity in cell shapes by estimating the tessellation parameters for each cell from the sparse data-points on its boundaries. We have tested the proposed 3D reconstruction method on time-lapse CLSM image stacks of the Arabidopsis Shoot Apical Meristem (SAM) and have shown that the AQVT based reconstruction method can correctly estimate the 3D shapes of a large number of SAM cells.
Highlights
The causal relationship between cell growth patterns and gene expression dynamics has been a major topic of interest in developmental biology
In Figure 8(A), we have shown the computationally resliced cell slices at various depths for Euclidean distance based Voronoi tessellation and Figure 8(B) shows the 2D cross sections for the same cells as obtained by reslicing the 3D cell shapes in the proposed Adaptive Quadratic Voronoi Tessellation’ (AQVT) based reconstruction method
We proposed a quadratic distance metric based Voronoi tessellation framework to capture the asymmetry of the cell sizes and growth along their different axes
Summary
The causal relationship between cell growth patterns and gene expression dynamics has been a major topic of interest in developmental biology. A proper quantitative analysis of the cell growth patterns in both the plant and the animal tissues has remained mostly elusive so far. Information such as rates and patterns of cell expansion play a critical role in explaining cell growth and deformation dynamics and thereby can be extremely useful in understanding morphogenesis. Confocal Laser Scanning Microscopy (CLSM) enables us to visually inspect the inner parts of the multilayered tissues. Through this technique we can image tissues as a collection of serial optical slices ( known as the ‘Z-Stack’), which can be used for analysis. Live cell imaging is a class of microscopy, where the same living cells are observed and imaged at regular time intervals over several hours to monitor their motion or displacement and to visualize cell growth and division dynamics
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