Original video: https://www.youtube.com/watch?v=evPCqmQ85YA
The process of visualizing the inner epidermal layer of Arabidopsis sepals involves detailed image processing rather than direct dissection, as dissecting flowers below stage 8 (103-238 µm) can damage the sepals. Using MorphoGraphX software, the inner epidermal layer is revealed by removing the underlying flower meristem voxels (a process referred to as 'voxel removal'). A longitudinal Clipping plane is positioned through the middle of the sepal, where curvature is minimal, allowing for clearer visualization of the inner cell layers.
To begin, the image is moved to the Work Stack for editing. By adjusting the Clipping plane depth to 20 µm or less, the sepal section can be optimized for minimal curvature. The flower meristem voxels below the inner epidermis are carefully removed using the 'Voxel Edit' tool, starting with a small radius (below 5 µm) for precise editing. Once the meristem voxels are removed, the tool's radius is increased to eliminate voxels from the remaining internal section, followed by smoothing out any unevenness.
This method provides a refined approach to revealing the internal sepal structure without damaging the sample, facilitating further analysis of growth and development in Arabidopsis sepals.
Keywords: growth, 2.5D segmentation, live imaging, image processing, deep tissue imaging, Arabidopsis, MorphoGraphX, sepals
Citation: Singh Yadav A and Roeder AHK (2024) An optimized live imaging and multiple cell layer growth analysis approach using Arabidopsis sepals. Front. Plant Sci. 15:1449195. doi: 10.3389/fpls.2024.1449195
Received: 14 June 2024; Accepted: 29 July 2024;
Published: 03 September 2024.
Attribution 4.0 International — CC BY 4.0 - Creative Commons
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