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Three-Dimensional Characterization of Cell Clusters Using Synchrotron-Radiation-Based Micro-Computed Tomography

Bert Müller, Marco Riedel and Philipp J. Thurner
Microscopy and Microanalysis
Vol. 12, No. 2, pp. 97-105, April 2006


Micro-computed tomography with the highly intense, monochromatic X-rays produced by the synchrotron is a superior method to nondestructively measure the local absorption in 3D space. Since biological tissues and cells consist mainly of water as the surrounding medium, higher absorbing agents have to be incorporated into the structures of interest. Even without X-ray optics, one can uncover the stain distribution with a spatial resolution of about 1 µm. Incorporating the stain at selected cell compartments, e.g. binding to the RNA/DNA, their density distribution becomes quantified. In this communication, we demonstrate that tomograms obtained at the beamlines BW2 and W2 (HASYLAB at DESY, Hamburg, Germany) and 4S (SLS, Villigen, Switzerland) clearly show that the RNA/DNA-stained cell clusters have a core of high density and a peripheral part of lower density, which correlate with results of optical microscopy. The inner part of the clusters is associated with non-vital cells as the result of insufficient oxygen and nutrition supply. This necrotic part is surrounded by (6 ± 1) layers of vital cells.

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  author = {Bert M\"uller and Marco Riedel and Philipp J. Thurner},
  title = {Three-Dimensional Characterization of Cell Clusters Using Synchrotron-Radiation-Based Micro-Computed Tomography},
  journal = {Microscopy and Microanalysis},
  year = {2006},
  month = {April},
  pages = {97-105},
  volume = {12},
  number = {2},
  keywords = {micro-computed tomography, synchrotron radiation, HEK 293 cell cluster, DNA/RNA staining, 3D characterization, segmentation}