title: Correlative light and volume electron microscopy: using focused ion beam scanning electron microscopy to image transient events in model organisms.
abstract: The study of a biological event within a live model organism has become routine through the use of fluorescent labeling of specific proteins in conjunction with laser confocal imaging. These methods allow 3D visualization of temporal events that can elucidate biological function but cannot resolve the tissue organization, extracellular and subcellular details of the tissues. Here, we present a method for correlating electron microscopy image data with the light microscopy data from the same sample volume to reveal the 3D structural information: "correlative light and volume electron microscopy." The methods for live video confocal microscopy, fixation and embedding of the tissue for electron microscopy, the focused ion beam scanning electron microscopy method for sequentially slicing and imaging the volume of interest, and the treatment of the resulting 3D dataset are presented. The method is illustrated with data collected during the angiogenesis of blood vessels in a transgenic zebrafish embryo.
authors.affiliation: The Nanovision Centre, School of Engineering and Materials Science, Queen Mary University of London, London, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Bushby
authors.forename: Giovanni
authors.initials: G
authors.lastname: Mariggi
authors.forename: Hannah E J
authors.initials: HE
authors.lastname: Armer
authors.forename: Lucy M
authors.initials: LM
authors.lastname: Collinson
mesh_terms: D000818:Animals; D001808:Blood Vessels / Q000648:ultrastructure; D055032:Electron Microscope Tomography; D004625:Embryo, Nonmammalian / Q000098:blood supply / Q000648:ultrastructure*; D004853:Epoxy Resins / Q000737:chemistry; D021621:Imaging, Three-Dimensional; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D008867:Microtomy; D018919:Neovascularization, Physiologic; D016613:Plastic Embedding; D012984:Software; D016707:Tissue Fixation; D015027:Zebrafish
chemical_list: D004853:Epoxy Resins; C025599:Durcupan ACM