Abstract
Callose (β-1,3-glucan) is both structural and functional component of plasmodesmata (Pd). The turnover of callose at Pd controls the cell-to-cell diffusion rate of molecules through Pd. An accurate assessment of changes in levels of Pd-associated callose has become a first-choice experimental approach in the research of intercellular communication in plants.
Here we describe a detailed and easy-to-perform procedure for imaging and quantification of Pd-associated callose using fixed plant tissue stained with aniline blue. We also introduce an automated image analysis protocol for non-biased quantification of callose levels at Pd from fluorescence images using ImageJ. Two experimental examples of Pd-callose quantification using the automated method are provided as well.
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Acknowledgement
We thank Eran Bosis at Tel Aviv University (TAU) for editing the ImageJ macro script. This work was supported by the Israel Science Foundation grant 723/00-17.1 to BLE, and by the Manna Institute for Plant Biosciences at TAU. RZ is partially supported by the Constantiner Institute for Molecular Genetics at TAU.
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Zavaliev, R., Epel, B.L. (2015). Imaging Callose at Plasmodesmata Using Aniline Blue: Quantitative Confocal Microscopy. In: Heinlein, M. (eds) Plasmodesmata. Methods in Molecular Biology, vol 1217. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1523-1_7
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DOI: https://doi.org/10.1007/978-1-4939-1523-1_7
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1522-4
Online ISBN: 978-1-4939-1523-1
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