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Array tomography: characterizing FAC‐sorted populations of zebrafish immune cells by their 3D ultrastructure
Author(s) -
WACKER IRENE,
CHOCKLEY PETER,
BARTELS CAROLIN,
SPOMER WALDEMAR,
HOFMANN ANDREAS,
GENGENBACH ULRICH,
SINGH SACHIN,
THALER MARLENE,
GRABHER CLEMENS,
SCHRÖDER RASMUS R.
Publication year - 2015
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12223
Subject(s) - ultrastructure , zebrafish , immune system , tomography , biology , microbiology and biotechnology , computational biology , evolutionary biology , anatomy , physics , immunology , genetics , gene , optics
Summary 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.

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