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Three-dimensional characterization of cell clusters using synchrotron-radiation-based micro-computed tomography.

Bert Müller1, Marco Riedel, Philipp J Thurner

  • 1Computer Vision Laboratory ETH Zürich, Gloriastrasse 35, CH-8092 Zürich, Switzerland. bmueller@vision.ee.ethz.ch

Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada
|May 8, 2007
PubMed
Summary

Synchrotron micro-computed tomography non-destructively visualizes cellular structures. RNA/DNA staining reveals necrotic cell cores within vital cell layers in HEK 293 clusters.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Medical Imaging

Background:

  • Biological tissues require contrast agents for X-ray absorption imaging.
  • Synchrotron micro-computed tomography (micro-CT) offers high-resolution 3D imaging.
  • Staining specific cellular components enhances contrast in micro-CT.

Purpose of the Study:

  • To demonstrate the utility of synchrotron micro-CT for visualizing cellular structures.
  • To quantify the distribution of RNA/DNA stains within cell clusters.
  • To correlate micro-CT findings with optical microscopy data.

Main Methods:

  • Utilized synchrotron micro-CT at beamlines BW2, W2, and 4S.
  • Employed RNA/DNA staining for contrast enhancement.
  • Analyzed tomograms of HEK 293 cell clusters.

Main Results:

  • Achieved spatial resolution of approximately 1 micrometer.
  • Observed distinct high-density core and lower-density periphery in stained cell clusters.
  • Correlated density variations with cell viability, identifying a necrotic core surrounded by vital cells.

Conclusions:

  • Synchrotron micro-CT effectively visualizes stain distribution in 3D at the cellular level.
  • The method can differentiate between vital and non-vital cells within clusters based on density.
  • Necrotic core formation in cell clusters is linked to oxygen and nutrient deprivation.