Improved Methods for Preparing Transverse Sections and Unrolled Whole Mounts of Maize Leaf Primordia for Fluorescence and Confocal Imaging

Janlo M. Robil; Cedric David T. Cortez; Chloe Margalaux R. Villafuerte; Edgar Daniel C. Dela Pe&#241;a; Derreck O. De Leon; Roselle Joy R. Rioja; Paula McSteen

doi:10.3791/65239

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Improved Methods for Preparing Transverse Sections and Unrolled Whole Mounts of Maize Leaf Primordia for Fluorescence and Confocal Imaging

JoVE Journal
Biology
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JoVE Journal Biology

Improved Methods for Preparing Transverse Sections and Unrolled Whole Mounts of Maize Leaf Primordia for Fluorescence and Confocal Imaging

2,653 Views

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06:11 min

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September 22, 2023

DOI:

10.3791/65239-v

DOI:

10.3791/65239-v

•

06:11 min

September 22, 2023

•

32 Views

Janlo M. Robil^1,2, Cedric David T. Cortez¹, Chloe Margalaux R. Villafuerte¹, Edgar Daniel C. Dela Peña¹, Derreck O. De Leon¹, Roselle Joy R. Rioja¹, Paula McSteen²

¹Department of Biology, School of Science and Engineering,Ateneo de Manila University, ²Division of Biological Sciences, Interdisciplinary Plant Group, and Missouri Maize Center,University of Missouri

Transcript

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.

Summary

Maize leaf primordia are deeply ensheathed and rolled, making them difficult to study. Here, we present methods for preparing transverse sections and unrolled whole mounts of maize leaf primordia for fluorescence and confocal imaging.