title: A 3D imaging and visualization workflow, using confocal microscopy and advanced image processing for brachyuran crab larvae.
abstract: Confocal laser scanning microscopy is an excellent tool for nondestructive imaging of arthropods and can provide detailed information on morphology including fine surface detail. A methodology is presented here for the visualization by confocal microscopy of arthropods, using brachyuran crab zoeal stages as examples and postprocessing techniques derived from micro-CT protocols to improve the final images. This protocol is divided into description of the preprocessing steps (cleaning, staining, digesting and mounting), confocal laser scanning microscopy and data visualization using open-source, freeware programs ImageJ and Drishti. The advantages of using ImageJ to standardize stack data and Drishti for surface rendering are discussed. The methodology has been comprehensively tested using data acquired from all four brands of confocal microscope (Leica, Nikon, Olympus and Zeiss).
authors.affiliation: School of Biological Sciences, Royal Holloway University of London, Egham, Surrey, UK.|Imaging and Analysis Centre, Core Research Laboratories, The Natural History Museum, London, UK.|Department of Life Sciences, The Natural History Museum, London, UK.
authors.forename: S A
authors.initials: SA
authors.lastname: Kamanli
authors.affiliation: German Centre for Marine Biodiversity Research, Senckenberg am Meer, Wilhelmshaven, Germany.
authors.forename: T C
authors.initials: TC
authors.lastname: Kihara
authors.affiliation: Imaging and Analysis Centre, Core Research Laboratories, The Natural History Museum, London, UK.
authors.forename: A D
authors.initials: AD
authors.lastname: Ball
authors.affiliation: School of Biological Sciences, Royal Holloway University of London, Egham, Surrey, UK.
authors.forename: D
authors.initials: D
authors.lastname: Morritt
authors.affiliation: Department of Life Sciences, The Natural History Museum, London, UK.
authors.forename: P F
authors.initials: PF
authors.lastname: Clark
mesh_terms: D000818:Animals; D001181:Arthropods / Q000033:anatomy & histology*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007814:Larva / Q000033:anatomy & histology; D018613:Microscopy, Confocal / Q000379:methods*; D057188:Workflow
keywords: 3D imaging; Brachyuran crab larvae; Drishti; ImageJ; confocal laser scanning microscopy; visualization