Light sheet fluorescence microscopy of plant roots growing on the surface of a gel Journal Article

Author(s): von Wangenheim, Daniel; Hauschild, Robert; Friml, Jiří
Article Title: Light sheet fluorescence microscopy of plant roots growing on the surface of a gel
Affiliation IST Austria
Abstract: One of the key questions in understanding plant development is how single cells behave in a larger context of the tissue. Therefore, it requires the observation of the whole organ with a high spatial- as well as temporal resolution over prolonged periods of time, which may cause photo-toxic effects. This protocol shows a plant sample preparation method for light-sheet microscopy, which is characterized by mounting the plant vertically on the surface of a gel. The plant is mounted in such a way that the roots are submerged in a liquid medium while the leaves remain in the air. In order to ensure photosynthetic activity of the plant, a custom-made lighting system illuminates the leaves. To keep the roots in darkness the water surface is covered with sheets of black plastic foil. This method allows long-term imaging of plant organ development in standardized conditions.
Keywords: Organogenesis; Arabidopsis; Fluorescence microscopy; Light sheet fluorescence microscopy; Light-sheet; Lightsheet microscopy; LSFM; Selective plane illumination microscopy; SPIM
Journal Title: Journal of visualized experiments JoVE
Volume: 2017
Issue 119
ISSN: 1940-087X
Publisher: Journal of Visualized Experiments  
Date Published: 2017-01-18
Start Page: Article number: e55044
Copyright Statement: CC BY-NC-ND
DOI: 10.3791/55044
Notes: We thank Matyáš Fendrych for critical reading/viewing and Stephan Stadlbauer for the audio equipment. Thanks to the Miba Machine Shop at IST Austria for their contribution to the OpenSPIM. The research leading to these results has received funding from the People Programme (Marie Curie Actions) of the European Union's Seventh Framework Programme (FP7/2007-2013) under REA grant agreement n° [291734] and European Research Council (project ERC-2011-StG-20101109-PSDP). ##BioimagingFacility #MaschineShop
Open access: yes (OA journal)