6533b7d8fe1ef96bd126a4dd
RESEARCH PRODUCT
Going beyond histology. Synchrotron micro-computed tomography as a methodology for biological tissue characterization: from tissue morphology to individual cells.
Helmut SchubertUlrich GrossUlrich GrossChristoph BrochhausenAstrid HaibelRolf ZehbeHeinrich RiesemeierC. James Kirkpatricksubject
Cartilage Articularthree-dimensional imagingMaterials scienceOpacityScanning electron microscope1004Biomedical EngineeringBiophysicsAnalytical chemistryBioengineeringPhoton energyIn Vitro TechniquesBiochemistrysynchrotron micro-computed tomographylaw.inventionBiomaterialshistologyChondrocyteslawConfocal microscopyResearch articlesAnimalscartilageCells CulturedTomographic reconstruction30HistologySynchrotron124Radiographic Image EnhancementTransmission electron microscopychondrocyteCattleTomography X-Ray ComputedSynchrotronsscanning electron microscopyBiotechnologyBiomedical engineeringdescription
Current light microscopic methods such as serial sectioning, confocal microscopy or multiphoton microscopy are severely limited in their ability to analyse rather opaque biological structures in three dimensions, while electron optical methods offer either a good three-dimensional topographic visualization (scanning electron microscopy) or high-resolution imaging of very thin samples (transmission electron microscopy). However, sample preparation commonly results in a significant alteration and the destruction of the three-dimensional integrity of the specimen. Depending on the selected photon energy, the interaction between X-rays and biological matter provides semi-transparency of the specimen, allowing penetration of even large specimens. Based on the projection-slice theorem, angular projections can be used for tomographic imaging. This method is well developed in medical and materials science for structure sizes down to several micrometres and is considered as being non-destructive. Achieving a spatial and structural resolution that is sufficient for the imaging of cells inside biological tissues is difficult due to several experimental conditions. A major problem that cannot be resolved with conventional X-ray sources are the low differences in density and absorption contrast of cells and the surrounding tissue. Therefore, X-ray monochromatization coupled with a sufficiently high photon flux and coherent beam properties are key requirements and currently only possible with synchrotron-produced X-rays. In this study, we report on the three-dimensional morphological characterization of articular cartilage using synchrotron-generated X-rays demonstrating the spatial distribution of single cells inside the tissue and their quantification, while comparing our findings to conventional histological techniques.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2009-03-28 | Journal of the Royal Society, Interface |