title: Light Sheet Fluorescence Microscopy of Plant Roots Growing on the Surface of a Gel.
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.
authors.affiliation: Developmental and Cell Biology of Plants, Institute of Science and Technology Austria; daniel.von.wangenheim@ist.ac.at.
authors.forename: Daniel
authors.initials: D
authors.lastname: von Wangenheim
authors.affiliation: Bioimaging Facility, Institute of Science and Technology Austria.
authors.forename: Robert
authors.initials: R
authors.lastname: Hauschild
authors.affiliation: Developmental and Cell Biology of Plants, Institute of Science and Technology Austria.
authors.forename: Jiří
authors.initials: J
authors.lastname: Friml
mesh_terms: D003624:Darkness; D005782:Gels; D008027:Light; D008856:Microscopy, Fluorescence / Q000379:methods*; D010788:Photosynthesis; D018515:Plant Leaves; D018517:Plant Roots / Q000254:growth & development*; D014867:Water
chemical_list: D005782:Gels; D014867:Water