Molecular imaging, emerging as an indispensable research tool, enhances the understanding of the physiological and pathological changes in organisms. This non-invasive technology is capable of visualizing the dynamic molecular and cellular processes taking place in vivo over a period of time, which has facilitated preclinical applications in biomedical in vivo studies, such as tumor detection at an early stage, stem cell imaging, and drug efficacy evaluation. Hybrid Imaging has recently become the new trend in the development of biomedical imaging. By combining different imaging modalities, it provides both anatomical structure and functional information, which brings together the merits of good sensitivity, high resolution, and fast speed. Hybrid imaging not only improves imaging quality, but also enables non-invasive in vivo 3D detection. This research focuses on reconstruction methods and prototype system of bioluminescence tomography based on hybrid imaging. It aims to solve biomedical problems in practical application with in-depth studies of theory and algorithms, as well as design of a hybrid imaging prototype system. In vivo experiments on small animals have been conducted to verify the feasibility of the proposed 3D reconstruction methods and the prototype system. The main contributions of this thesis include: 1.A parallel iterative shrinkage algorithm has been presented for non-invasive in vivo 3D detection of small liver tumors in the mouse. The existing 3D reconstruction algorithms have partly solved the problems of accuracy and speed, while most experiments were based on numerical simulation, geometric phantoms, or a mouse phantom. Since the signal is usually much weaker in practical application, it is necessary to validate the algorithms with in vivo experiments based on the orthotopic mouse tumor model. This method takes advantage of X-ray computed tomography and multi-view bioluminescence imaging, providing anatomical structure and bioluminescence intensity information to reconstruct the size and location of tumors. It guarantees the accuracy, efficiency and reliability for 3D reconstruction by incorporating some mathematical strategies including specific smooth convex approximation, iterative shrinkage operator and affine subspace. Finally, an in vivo experiment on an HCCLM3 orthotopic xenograft mouse model has been conducted. The findings indicate that a tiny lesion can be localized with a position bias no more than 1mm, and the co...
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