Visualization of the 3D cell distribution in biological tissues using contrast-enhanced microfocus computed tomography – an optimization study.

Details

Supervisors

Greet KERCKHOFS

Faculty

Faculté de pharmacie et des sciences biomédicales

Degree label

Master [120] en sciences biomédicales, à finalité approfondie

Abstract

This study investigates the optimization of Contrast-Enhanced Computed Tomography (CECT) imaging for soft tissues using CA4+. Tissue visualization is essential for research. Currently, the reference technique used to visualize the composition and organization of tissue microstructure is 2D histology. Another imaging method is the use of microfocus X-ray computed tomography (Micro-CT), which enables 3D imaging. Murine ears were selected as the experimental model seeing their diverse tissue composition, offering a large basis to evaluate CA4+ specificity and imaging protocols. Building upon Dr. Hoffmann's work, this research explores the reproducibility of CA4+ staining and its ability to highlight cellular nuclei. The project deals with challenges such as motion artifacts and resolution variability. A new pipeline was developed, covering staining, sample mounting, image acquisition, reconstruction and segmentation techniques, including deep learning. By comparing CA4+ staining with traditional histological methods, such as Toluidine Blue staining, the study validated the agent's specificity while identifying its limitations and broader applications. Significant advancements were achieved in reducing imaging time without compromising data quality. A cooling stage has been implemented to mitigate heat-induced motion artifacts, enhancing stability during prolonged imaging sessions. Advanced segmentation approaches were employed to extract 3D volumetric data, revealing insights into the 3D distribution of cell nuclei. These findings feature the potential of CA4+ and micro-CT to improve high-resolution, non-destructive 3D imaging of soft tissues.