title: Array tomography: characterizing FAC-sorted populations of zebrafish immune cells by their 3D ultrastructure.
abstract: For 3D reconstructions of whole immune cells from zebrafish, isolated from adult animals by FAC-sorting we employed array tomography on hundreds of serial sections deposited on silicon wafers. Image stacks were either recorded manually or automatically with the newly released ZEISS Atlas 5 Array Tomography platform on a Zeiss FEGSEM. To characterize different populations of immune cells, organelle inventories were created by segmenting individual cells. In addition, arrays were used for quantification of cell populations with respect to the various cell types they contained. The detection of immunological synapses in cocultures of cell populations from thymus or WKM with cancer cells helped to identify the cytotoxic nature of these cells. Our results demonstrate the practicality and benefit of AT for high-throughput ultrastructural imaging of substantial volumes.
authors.affiliation: Centre for Advanced Materials, Universität Heidelberg, Heidelberg, Germany.|Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.
authors.forename: Irene
authors.initials: I
authors.lastname: Wacker
authors.affiliation: Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Chockley
authors.affiliation: Centre for Advanced Materials, Universität Heidelberg, Heidelberg, Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Bartels
authors.affiliation: Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.|Institute for Applied Computer Science, Karlsruhe Institute of Technology, Karlsruhe, Germany.
authors.forename: Waldemar
authors.initials: W
authors.lastname: Spomer
authors.affiliation: Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.|Institute for Applied Computer Science, Karlsruhe Institute of Technology, Karlsruhe, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hofmann
authors.affiliation: Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.|Institute for Applied Computer Science, Karlsruhe Institute of Technology, Karlsruhe, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Gengenbach
authors.affiliation: Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany.
authors.forename: Sachin
authors.initials: S
authors.lastname: Singh
authors.affiliation: Carl Zeiss Microscopy GmbH, Oberkochen, Germany.
authors.forename: Marlene
authors.initials: M
authors.lastname: Thaler
authors.affiliation: Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.|Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany.
authors.forename: Clemens
authors.initials: C
authors.lastname: Grabher
authors.affiliation: Centre for Advanced Materials, Universität Heidelberg, Heidelberg, Germany.|Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.|Cryo-EM, CellNetworks, BioQuant Universitätsklinikum Heidelberg, Heidelberg, Germany.
authors.forename: Rasmus R
authors.initials: RR
authors.lastname: Schröder
mesh_terms: D000328:Adult; D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007107:Immune System / Q000166:cytology* / Q000648:ultrastructure*; D054992:Immunological Synapses / Q000648:ultrastructure; D008214:Lymphocytes / Q000648:ultrastructure*; D013950:Thymus Gland / Q000166:cytology / Q000648:ultrastructure; D014054:Tomography / Q000379:methods*; D015027:Zebrafish
keywords: 3D reconstruction; array tomography; cytotoxic cells; immunological synapse; large volume ultrastructure; zebrafish