ZFIN ID: ZDB-PUB-150123-4
Array tomography: characterizing FAC-sorted populations of zebrafish immune cells by their 3D ultrastructure
Wacker, I., Chockley, P., Bartels, C., Spomer, W., Hofmann, A., Gengenbach, U., Singh, S., Thaler, M., Grabher, C., Schröder, R.R.
Date: 2015
Source: Journal of microscopy   259(2): 105-13 (Journal)
Registered Authors: Grabher, Clemens, Singh, Sachin
Keywords: 3D reconstruction, array tomography, cytotoxic cells, immunological synapse, large volume ultrastructure, zebrafish
MeSH Terms:
  • Adult
  • Animals
  • Cell Line, Tumor
  • Cell Movement
  • Cell Separation
  • Cells, Cultured
  • Flow Cytometry/methods
  • Humans
  • Imaging, Three-Dimensional/methods*
  • Immune System/cytology*
  • Immune System/ultrastructure*
  • Immunological Synapses/ultrastructure
  • Lymphocytes/ultrastructure*
  • Thymus Gland/cytology
  • Thymus Gland/ultrastructure
  • Tomography/methods*
  • Zebrafish
PubMed: 25611576 Full text @ J. Micros.
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.
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