Grasses like maize have leaf primordia grow within the shoot, making them difficult to study. We address this problem with improved protocols for preparing maize leaf transverse sections and whole mounts for fluorescence and confocal imaging. The first protocol uses a wire stripper to cut the older leaves, allowing the measurement of the primordium prior to sectioning.
The second protocol uses clear, double-sided nano tape to unroll whole leaf primordium. These protocols improve the accuracy of transverse sectioning and enable unrolling of the leaf primordia, which have been very difficult to achieve due to their rolled, conical morphology. These methods will be useful for visualizing and quantifying leaf anatomical and developmental traits in maize and other grasses.
Based on these methods, we plan to develop live cell imaging strategies to address questions in grass leaf development.