TY - JOUR
T1 - Tomography studies of human foreskin fibroblasts on polymer yarns
AU - Thurner, Philipp
AU - Müller, Bert
AU - Beckmann, Felix
AU - Weitkamp, Timm
AU - Rau, Christoph
AU - Müller, Ralph
AU - Hubbell, Jeffrey A.
AU - Sennhauser, Urs
N1 - Funding Information:
The authors thank Martin Müller of ETH Zürich for SEM sample preparation. The authors gratefully acknowledge financial support from the Swiss National Science Foundation (2153-057127.99), HASYLAB/DESY II-99-077: ‘Computed Microtomography’, and ESRF LS-1980: ‘3D Imaging of Biological Cells and Scaffolds using X-ray Microtomography’.
PY - 2003/1
Y1 - 2003/1
N2 - Cell culture experiments are usually performed as in vitro studies based on 2D seeding and characterization (light microscopy). With respect to the in vivo situation, however, 2D studies are often inappropriate due to the 3D character of living tissue in nature. Textiles with their versatile 3D structures are chosen as suitable scaffolds in tissue engineering for 3D in vitro studies. Micro-computed tomography using X-rays (μCT) belongs to the most promising techniques for isotropic, noninvasive 3D characterization. Using synchrotron radiation (SRμCT) the spatial resolution can be extended to the sub-micrometer range well below cell size. μCT does not need vacuum conditions making experiments in the hydrated state possible, as we show by data from SRμCT acquired at second and third-generation synchrotron sources. We seeded human foreskin fibroblasts on polymer multifilament yarns. These composites, embedded in a hydrogel or fluid, are held in thin-walled glass capillaries. Since the composites consist of light elements, the cells have to be labeled for visualization by the use of highly absorptive agents, osmium and gold. In order to hold the label concentration as low as possible, we present a way to choose the photon energy for which the minimum concentration is reached. Differences in threshold selection for second- and third-generation synchrotron sources are pointed out, revealing the advantages of both types with respect to quantitative analysis. The study is based on appropriate staining methods and protocols developed in our laboratory. With the results we demonstrate that SRμCT yields images similar to established electron and light microscopy but uncovers also the microstructure in 3D space.
AB - Cell culture experiments are usually performed as in vitro studies based on 2D seeding and characterization (light microscopy). With respect to the in vivo situation, however, 2D studies are often inappropriate due to the 3D character of living tissue in nature. Textiles with their versatile 3D structures are chosen as suitable scaffolds in tissue engineering for 3D in vitro studies. Micro-computed tomography using X-rays (μCT) belongs to the most promising techniques for isotropic, noninvasive 3D characterization. Using synchrotron radiation (SRμCT) the spatial resolution can be extended to the sub-micrometer range well below cell size. μCT does not need vacuum conditions making experiments in the hydrated state possible, as we show by data from SRμCT acquired at second and third-generation synchrotron sources. We seeded human foreskin fibroblasts on polymer multifilament yarns. These composites, embedded in a hydrogel or fluid, are held in thin-walled glass capillaries. Since the composites consist of light elements, the cells have to be labeled for visualization by the use of highly absorptive agents, osmium and gold. In order to hold the label concentration as low as possible, we present a way to choose the photon energy for which the minimum concentration is reached. Differences in threshold selection for second- and third-generation synchrotron sources are pointed out, revealing the advantages of both types with respect to quantitative analysis. The study is based on appropriate staining methods and protocols developed in our laboratory. With the results we demonstrate that SRμCT yields images similar to established electron and light microscopy but uncovers also the microstructure in 3D space.
KW - Biological physics spectroscopic- and microscopic techniques
KW - Cell complexes (geometry and topology)
KW - Computerized tomography
KW - Image processing algorithms
KW - Synchrotron radiation
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U2 - 10.1016/S0168-583X(02)01729-9
DO - 10.1016/S0168-583X(02)01729-9
M3 - Article
AN - SCOPUS:0037243843
VL - 200
SP - 397
EP - 405
JO - Nuclear Instruments and Methods in Physics Research, Section B: Beam Interactions with Materials and Atoms
JF - Nuclear Instruments and Methods in Physics Research, Section B: Beam Interactions with Materials and Atoms
SN - 0168-583X
ER -