Abstract
Complex geometry of plant organs and various types of organ surface deformation, including growth or hygroscopic movements, can be analyzed using sequential replica method. It enables obtaining a time-lapse series of high resolution images visualizing details of the examined surface and provides data sufficient for detailed computation of parameters characterizing surface deformation and geometry. Series of molds, made in dental polymer, representing the examined surface are used to obtain casts in epoxy resin or nail polish replicas, which are ready for microscopic examination, while the structure itself remains intact. Images obtained from the epoxy casts in scanning electron microscopy can be further used for 3D reconstruction and computation of local geometry. The sequential replica method is a universal method and can be applied to image complex shapes of a range of structures, like meristems, flowers, leaves, scarious bracts, or trichomes. Different plant species growing in various conditions can be studied.
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Acknowledgments
The sequential replica method has been developed by the late Paul B. Green and writing this chapter we have used his numerous indispensable advices that we have learnt from Dr. Jacques Dumais, the last graduate student of Paul. We would like to thank Dr. Ewa Teper (Laboratory of Scanning Electron Microscopy, Faculty of Earth Sciences, University of Silesia) for help with scanning electron microscopy, and Drs. Joanna Elsner and Aleksandra Rypień (Faculty of Biology and Environment Protection, University of Silesia) for providing SEM micrographs of leaf and bract epidermis, respectively. This work in D.K. lab was financially supported by the MAESTRO research grant No 2011/02/A/NZ3/00079 (for D.K) and the SONATA BIS6 research grant No 2016/22/E/NZ3/00342 (for A.B.) both from the National Science Centre, Poland.
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Kwiatkowska, D., Natonik-Białoń, S., Burian, A. (2019). Sequential Replicas: Method for In Vivo Imaging of Plant Organ Surfaces that Undergo Deformation. In: Cvrčková, F., Žárský, V. (eds) Plant Cell Morphogenesis. Methods in Molecular Biology, vol 1992. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9469-4_16
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DOI: https://doi.org/10.1007/978-1-4939-9469-4_16
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