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Browsing Senior Theses & Projects by Author "Adams, Ellen"

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    Selective Plane Illumination Microscopy (SPIM) for Hydra Development
    (2021) Adams, Ellen; Collins, Eva-Maria; Delano, Maggie
    A challenge in biology is imaging live biological samples in 3D to capture larger-scale dynamic processes. These include whole-organism processes like development and regeneration, which involve dramatic rearrangements of cells and changes in cell chape. To image these morphological changes in biological samples, transgenic animals can be generated that have fluorescent proteins to label subcellular structures, cells, or tissues (Takeuchi et al., 1999; Wittlieb et al., 2006). This fluorescence allows for the visualization of the labelled sample and can be used to track cells and cell contents, as well as shape changes (Wittlieb et al., 2006; Hadjantonakis et al., 2004). One of the difficulties of imaging these fluorescent live samples is achieving good resolution in live samples due to the size of the specimen and the scattering properties of intact tissue (Huisken and Stainier, 2009). Due to these challenges, often live samples will instead be fixed and sectioned at various timepoints to achieve better visualization, however this approach does not lend itself to studying the continuous timeline of development. Selective Plane Illumination Microscopy (SPIM) allows for optical, instead of physical, sectioning for deeper penetration into the sample, to image live, intact embryos in 3D over time (Huisken and Stainier, 2009). SPIM has been used to study the development of embryos of the polyclad flatworm M. crozieri and fruit fly Drosophilia melanogaster (Girstmair et al., 2016; Schmied and Tomancak, 2016), as well as zebrafish embryo vasculature development over the course of up to five days (Daetwyler et al., 2019). In these studies, SPIM proved to be a useful tool for longer term continuous imaging of development in live embryos. We want to use this microscopy technique to visualize the development of Hydra, which are small freshwater cnidarians famous for their regenerative abilities. They regenerate from a small tissue piece or cell aggregate, and we aim to track cell movements throughout this process to follow the patterning of tissues that occurs. By applying the SPIM technique to studying regeneration and development in Hydra, we can gain insights into the dynamics of development and regeneration to gain a greater understanding of them.