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Three-dimensional characterization of HEK293 cell clusters using synchrotron-radiation-based micro-computed tomography

Three-dimensional characterization of HEK293 cell clusters using synchrotron-radiation-based micro-computed tomography
Three-dimensional characterization of HEK293 cell clusters using synchrotron-radiation-based micro-computed tomography
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 three-dimensional space. Because 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 such as refractive lens, one can uncover the stain distribution with the spatial resolution of about 1 [mu]m. Incorporating the stain at selected cell compartments, for example, 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 HEK 293 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 nonvital cells as the result of insufficient oxygen and nutrition supply. This necrotic part is surrounded by (6 ± 1) layers of vital cells.
micro-computed tomography, synchrotron radiation, HEK 293 cell cluster, DNA/RNA staining, 3D characterization, segmentation
1431-9276
97-105
Müller, Bert
acba4294-b684-4a09-81ac-32de31d39923
Riedel, Marco
1cfbb6b5-955a-40ce-bd61-894b5b76266a
Thurner, Philipp J.
ab711ddd-784e-48de-aaad-f56aec40f84f
Müller, Bert
acba4294-b684-4a09-81ac-32de31d39923
Riedel, Marco
1cfbb6b5-955a-40ce-bd61-894b5b76266a
Thurner, Philipp J.
ab711ddd-784e-48de-aaad-f56aec40f84f

Müller, Bert, Riedel, Marco and Thurner, Philipp J. (2006) Three-dimensional characterization of HEK293 cell clusters using synchrotron-radiation-based micro-computed tomography. Microscopy and Microanalysis, 12 (2), 97-105. (doi:10.1017/S1431927606060168).

Record type: Article

Abstract

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 three-dimensional space. Because 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 such as refractive lens, one can uncover the stain distribution with the spatial resolution of about 1 [mu]m. Incorporating the stain at selected cell compartments, for example, 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 HEK 293 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 nonvital 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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More information

Published date: April 2006
Keywords: micro-computed tomography, synchrotron radiation, HEK 293 cell cluster, DNA/RNA staining, 3D characterization, segmentation

Identifiers

Local EPrints ID: 48829
URI: http://eprints.soton.ac.uk/id/eprint/48829
ISSN: 1431-9276
PURE UUID: 341770d4-2785-4ab6-919d-fb18dadabe1e
ORCID for Philipp J. Thurner: ORCID iD orcid.org/0000-0001-7588-9041

Catalogue record

Date deposited: 16 Oct 2007
Last modified: 15 Mar 2024 09:50

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Contributors

Author: Bert Müller
Author: Marco Riedel

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