Abstract
Confocal microscopy is a technique widely used to live-image plant tissue. Cells can be visualized by using fluorescent probes that mark the cell wall or plasma membrane. This enables the confocal microscope to be used as a 3D scanner with submicron precision. Here we present a protocol using the 3D image processing software MorphoGraphX (http://www.MorphoGraphX.org) to extract the surface geometry and cell shapes in the shoot apex. By segmenting cells over consecutive time points, precise growth maps of the shoot apex can be produced. It is also possible to tag a protein of interest with a fluorescent marker and quantify protein expression at the cellular level.
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References
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Acknowledgments
We gratefully acknowledge all those involved in the development of the MorphoGraphX—Naomi Nakayama, Thierry Schuepbach, Alain Weber, Anne-Lise Routier-Kierzkowska, and Micha Hersch. We thank Daniel Kierzkowski for time series images and Cris Kuhlemeier for discussions. This work was supported by SystemX.ch, the Swiss National Science Foundation, EMBO Long-Term Fellowship to S.R., and the University of Bern.
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de Reuille, P.B., Robinson, S., Smith, R.S. (2014). Quantifying Cell Shape and Gene Expression in the Shoot Apical Meristem Using MorphoGraphX. In: Žárský, V., Cvrčková, F. (eds) Plant Cell Morphogenesis. Methods in Molecular Biology, vol 1080. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-643-6_10
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DOI: https://doi.org/10.1007/978-1-62703-643-6_10
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-642-9
Online ISBN: 978-1-62703-643-6
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