3D measurement of cell thickness and its dynamics in plants.

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conclusionA novel and efficient method was developed to accurately measure thickness in 3D of many cells from a confocal stack, as well as to track changes in cell thickness overtime. Plant cells and organs are three-dimensional objects with a certain thickness. Among basic geometric parameters (length, width, depth/thickness), cell thickness is less accurately and comprehensively measured, probably because it cannot be directly seen. The current methods of cell thickness quantification have some limitations, such as measuring only from a cross-section, not accounting for the directionality of biological thickness, or not offering a way to track changes in thickness of individual cells over time. This research is an attempt to bridge the gap, by making the quantification of thickness and tracking its changes in many cells easier and more accurate. We devised a novel method to efficiently measure average cell thickness in 3D from cells imaged with confocal microscopy, the most popular technique to image live samples over many days. The method, in combination with the popular software MorphoGraphX, also allows accurate and efficient tracking of changes in thickness between different time points. We tested the method on various organs of the model plant Arabidopsis thaliana such as the shoot apical meristem, the hypocotyl, the cotyledon, and the sepal. We demonstrated that this new method can reliably measure the thickness of hundreds of cells at once in a short amount of time to reveal new biological insights. We believe this would be a useful tool for plant researchers to accurately characterize this hidden morphological dimension.