Ms. Catherine Anne Heyward Profile

Catherine Anne Heyward

at Univ of Oslo

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SPIE Journal Paper | 22 November 2024 Open Access

Advanced soft tissue visualization in conjunction with bone structures using contrast-enhanced micro-CT

Torben Hildebrand, Qianli Ma, Catherine A. Heyward, Håvard Haugen, Liebert Nogueira

JMI, Vol. 11, Issue 06, 066001, (November 2024) https://doi.org/10.1117/12.10.1117/1.JMI.11.6.066001

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PurposeMicro-computed tomography (CT) analysis of soft tissues alongside bone remains challenging due to significant differences in X-ray absorption, preventing spatial inspection of bone remodeling including the cellular intricacies of mineralized tissues in developmental biology and pathology. The goal was to develop a protocol for contrast-enhanced micro-CT imaging that effectively visualizes soft tissues and cells in conjunction with bone while minimizing bone attenuation by decalcification.ApproachMurine femur samples were decalcified in ethylenediaminetetraacetic acid and treated with three different contrast agents: (i) iodine in ethanol, (ii) phosphotungstic acid in water, and (iii) Lugol’s iodine. Micro-CT scans were performed in the laboratory setup SkyScan 1172 and at the synchrotron radiation for medical physics beamline in synchrotron radiation facility Elettra. Soft and hard tissue contrast-to-noise ratio (CNR) and contrast efficiency after decalcification were measured.ResultsIn laboratory micro-CT, Lugol’s iodine demonstrated a threefold higher CNR in the bone marrow, representing the soft tissue portion, compared with the bone. Contrast efficiencies, measured in synchrotron micro-CT, were consistent with these findings. Higher resolutions and the specificity of Lugol’s iodine to cellular structures enabled detailed visualization of bone-forming cells in the epiphyseal plate.ConclusionsThe combination of decalcification and the utilization of the contrast agent Lugol’s iodine facilitated an enhanced soft tissue visualization in conjunction with bone.

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Proceedings Article | 17 October 2024 Presentation + Paper

Advanced soft tissue visualization in conjunction with bone structures using contrast-enhanced micro-CT

Torben Hildebrand, Qianli Ma, Catherine Anne Heyward, Håvard Haugen, Liebert Nogueira

Proceedings Volume 13152, 131520J (2024) https://doi.org/10.1117/12.3027063

KEYWORDS: Bone, Tissues, Iodine, Contrast agents, Biological samples, Visualization, Biological imaging, Synchrotrons, Cartilage

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Micro-CT analysis is conventionally used to investigate mineralized tissues. However, the potential of phase-contrast and contrast-enhanced micro-CT extends to soft tissue inspection, though optimizing soft tissue contrast alongside bone remains challenging, preventing spatial inspection of bone remodeling including the cellular components. The goal was to develop a protocol for contrast-enhanced micro-CT imaging that effectively visualizes soft tissues and cells in conjunction with bone while minimizing bone attenuation by decalcification. Murine femur samples were decalcified in ethylenediaminetetraacetic acid (EDTA) and treated with three different contrast agents: i) iodine in ethanol, ii) phosphotungstic acid (PTA) in water and iii) Lugol’s iodine. Micro-CT scans were performed in the laboratory set-up SkyScan 1172 and at the SYRMEP beamline in ELETTRA. Soft- and hard-tissue contrast-to-noise ratio (CNR) and contrast efficiency after decalcification were measured. In laboratory micro-CT, iodine in ethanol and PTA provided a higher CNR for bone compared to bone marrow, while Lugol’s iodine demonstrated a three-times higher CNR in bone marrow, representing the soft tissue portion. Contrast efficiencies, measured as the percentage increase of gray value compared to its decalcified state prior to contrast enhancement, were consistent with these findings in synchrotron micro-CT. Higher resolutions and the specificity of Lugol’s iodine to cellular structures enabled detailed visualization of bone-forming cells in the epiphyseal plate. The optimized protocol for micro-CT imaging significantly enhances soft tissue visualization alongside bone, facilitating non-invasive anatomical and histological analysis. This approach shows high potential for exploring the cellular intricacies of mineralized tissues in developmental biology and pathology.

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