title: Preparation of plants for developmental and cellular imaging by light-sheet microscopy.
abstract: Long-term fluorescence live-cell imaging experiments have long been limited by the effects of excitation-induced phototoxicity. The advent of light-sheet microscopy now allows users to overcome this limitation by restricting excitation to a narrow illumination plane. In addition, light-sheet imaging allows for high-speed image acquisition with uniform illumination of samples composed of multiple cell layers. The majority of studies conducted thus far have used custom-built platforms with specialized hardware and software, along with specific sample handling approaches. The first versatile commercially available light-sheet microscope, Lightsheet Z.1, offers a number of innovative solutions, but it requires specific strategies for sample handling during long-term imaging experiments. There are currently no standard procedures describing the preparation of plant specimens for imaging with the Lightsheet Z.1. Here we describe a detailed protocol to prepare plant specimens for light-sheet microscopy, in which Arabidopsis seeds or seedlings are placed in solid medium within glass capillaries or fluorinated ethylene propylene tubes. Preparation of plant material for imaging may be completed within one working day.
authors.affiliation: Department of Cell Biology, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Ovečka
authors.affiliation: Department of Cell Biology, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Vaškebová
authors.affiliation: Department of Cell Biology, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: George
authors.initials: G
authors.lastname: Komis
authors.affiliation: Department of Cell Biology, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Ivan
authors.initials: I
authors.lastname: Luptovčiak
authors.affiliation: Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA.
authors.forename: Andrei
authors.initials: A
authors.lastname: Smertenko
authors.affiliation: Department of Cell Biology, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Jozef
authors.initials: J
authors.lastname: Šamaj
mesh_terms: D017360:Arabidopsis; D008856:Microscopy, Fluorescence; D059828:Plant Cells; D063245:Plant Development; D016610:Tissue Embedding / Q000379:methods*