Array tomography: characterizing FAC-sorted populations of zebrafish immune cells by their 3D ultrastructure.

Autor: Wacker I; Centre for Advanced Materials, Universität Heidelberg, Heidelberg, Germany.; Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany., Chockley P; Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany., Bartels C; Centre for Advanced Materials, Universität Heidelberg, Heidelberg, Germany., Spomer W; Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.; Institute for Applied Computer Science, Karlsruhe Institute of Technology, Karlsruhe, Germany., Hofmann A; Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.; Institute for Applied Computer Science, Karlsruhe Institute of Technology, Karlsruhe, Germany., Gengenbach U; Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.; Institute for Applied Computer Science, Karlsruhe Institute of Technology, Karlsruhe, Germany., Singh S; Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany., Thaler M; Carl Zeiss Microscopy GmbH, Oberkochen, Germany., Grabher C; Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.; Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany., Schröder RR; Centre for Advanced Materials, Universität Heidelberg, Heidelberg, Germany.; Heidelberg Karlsruhe Research Partnership, Heidelberg/Karlsruhe, Germany.; Cryo-EM, CellNetworks, BioQuant Universitätsklinikum Heidelberg, Heidelberg, Germany.
Jazyk: angličtina
Zdroj: Journal of microscopy [J Microsc] 2015 Aug; Vol. 259 (2), pp. 105-113. Date of Electronic Publication: 2015 Jan 21.
DOI: 10.1111/jmi.12223
Abstrakt: 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.
(© 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.)
Databáze: MEDLINE
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