title: A customized light sheet microscope to measure spatio-temporal protein dynamics in small model organisms.
abstract: We describe a customizable and cost-effective light sheet microscopy (LSM) platform for rapid three-dimensional imaging of protein dynamics in small model organisms. The system is designed for high acquisition speeds and enables extended time-lapse in vivo experiments when using fluorescently labeled specimens. We demonstrate the capability of the setup to monitor gene expression and protein localization during ageing and upon starvation stress in longitudinal studies in individual or small groups of adult Caenorhabditis elegans nematodes. The system is equipped to readily perform fluorescence recovery after photobleaching (FRAP), which allows monitoring protein recovery and distribution under low photobleaching conditions. Our imaging platform is designed to easily switch between light sheet microscopy and optical projection tomography (OPT) modalities. The setup permits monitoring of spatio-temporal expression and localization of ageing biomarkers of subcellular size and can be conveniently adapted to image a wide range of small model organisms and tissue samples.
authors.affiliation: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Heraklion, Crete, Greece.
authors.forename: Matthias
authors.initials: M
authors.lastname: Rieckher
authors.affiliation: Institute of Electronic Structure and Laser, Foundation for Research and Technology-Hellas, Heraklion, Crete, Greece.
authors.forename: Ilias
authors.initials: I
authors.lastname: Kyparissidis-Kokkinidis
authors.affiliation: Institute of Electronic Structure and Laser, Foundation for Research and Technology-Hellas, Heraklion, Crete, Greece.
authors.forename: Athanasios
authors.initials: A
authors.lastname: Zacharopoulos
authors.affiliation: Institute of Electronic Structure and Laser, Foundation for Research and Technology-Hellas, Heraklion, Crete, Greece.
authors.forename: Georgios
authors.initials: G
authors.lastname: Kourmoulakis
authors.affiliation: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Heraklion, Crete, Greece; Department of Basic Sciences, Faculty of Medicine, University of Crete, Heraklion, Crete, Greece.
authors.forename: Nektarios
authors.initials: N
authors.lastname: Tavernarakis
authors.affiliation: Department of Bioengineering and Aerospace Engineering, Universidad Carlos III de Madrid, Madrid, Spain; Experimental Medicine and Surgery Unit, Instituto de Investigación Sanitaria del Hospital Gregorio Marañón, Madrid, Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Ripoll
authors.affiliation: Institute of Electronic Structure and Laser, Foundation for Research and Technology-Hellas, Heraklion, Crete, Greece.
authors.forename: Giannis
authors.initials: G
authors.lastname: Zacharakis
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism; D039642:Eukaryotic Initiation Factors / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*
chemical_list: D029742:Caenorhabditis elegans Proteins; D039642:Eukaryotic Initiation Factors; D049452:Green Fluorescent Proteins