Development of new methods for histology and fluorescence imaging through the exploration of potential symplastic domains in the endosperm of mature seeds

Diplôme Master Development of new methods for histology and fluorescence imaging through the exploration of potential symplastic domains in the endosperm of mature seeds eng 2024-01-31 author Schmitt Alexandra 898803 director Loubery Sylvain Philippe 0000-0001-7061-9456 669207 Département des sciences végétales (BIVEG) director Lopez Molina Luis 0000-0003-0463-1187 28870 Département des sciences végétales (BIVEG) director Sadian Yashar 917818 65 The objective of this Master’s project was to develop new methods for plant histology and fluorescence imaging, useful to the Department of Plant Sciences at Geneva University (UNIGE). Seeds were chosen as a biological sample as they are challenging; a successful protocol could thus easily be adapted to other plant material. A biological inquiry was also investigated and the existence of potential symplastic domains in the endosperm of mature seeds was probed. In this context, two transgenic lines were generated: a sequestered (pABI5::NLS-2mVenus) and a freely diffusible reporter line (pABI5::mVenus). Comparing expression domains of mVenus in both lines will reveal how the endosperm’s chalazal, micropylar and peripheral regions are symplastically connected. The first methods investigated were High-pressure freezing and Freeze substitution, followed by in situ Correlative light and electron microscopy. Secondly, fluorescence microscopy was extensively explored, and different methods were experimented with, such as tert-Butanol-dehydration, Technovit 8100-embedding, clearing on section, immunofluorescence, serial sectioning, confocal and 2-Photon microscopy combined with Fluorescence lifetime imaging. Dewey 570 Molecular Plant Sciences (MPS) 30 Section de biologie 16 false 831 Groupe Lopez-Molina 1d1f9022-6c32-4f50-aaf9-f2dbe0c11330 Master thesis Restricted Master_Schmitt_2024 FINAL.pdf 1779923 application/pdf 2d02db9897fe9874b1d06902aa9e29efbce8f72bd08b2fab45f74c3db06d2057 restrictedAccess Faculté des sciences / Section de biologie

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Development of new methods for histology and fluorescence imaging through the exploration of potential symplastic domains in the endosperm of mature seeds				2024	83	10

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Schmitt, Alexandra

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