Light Sheet Fluorescence microscopy of plant roots growing on the surface of a gel Von Wangenheim, Daniel ; https://orcid.org/0000-0002-6862-1247 Hauschild, Robert ; https://orcid.org/0000-0001-9843-3522 Friml, Jirí ; https://orcid.org/0000-0002-8302-7596 ddc:580 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. The Video is licensed under a CC BY NC ND license. Institute of Science and Technology Austria 2017 info:eu-repo/semantics/other doc-type:ResearchData ResearchData http://purl.org/coar/resource_type/63NG-B465 https://research-explorer.ista.ac.at/record/5565 https://research-explorer.ista.ac.at/download/5565/5599 von Wangenheim D, Hauschild R, Friml J. Light Sheet Fluorescence microscopy of plant roots growing on the surface of a gel. 2017. doi:<a href="https://doi.org/10.15479/AT:ISTA:66">10.15479/AT:ISTA:66</a> info:eu-repo/semantics/altIdentifier/doi/10.15479/AT:ISTA:66 info:eu-repo/semantics/openAccess