Significance: The multimodality imaging system has become a powerful tool for in-vivo biomedical research. However, a conventional multimodality system generally employs two independent detectors, which is costly and bulky. Meanwhile, the geometric cocalibration and image registration between the imaging modalities are also complicated.

Aim: To acquire the multimodality images for small animals with only one visible light sensed single-pixel detector.

Approach: The system is built based on a structured detection Fourier single-pixel imaging architecture. A cesium iodide doped with thallium [CsI(Tl)] scintillator plate is placed behind the sample in x-ray imaging, so the x-ray images can be converted to be visible and sensed with the same single-pixel detector as applied in fluorescence imaging.

Results: The spatial resolution of x-ray imaging was measured to be 1.81 mm, the sensitivity and the imaging depth of fluorescence imaging was evaluated to be ∼1.48 nmol / ml and 4 mm, respectively. In vivo multimodality imaging of a C57BL/6 female mouse bearing tumor targeted with mCherry was carried out.

Conclusions: We proposed an x-ray and fluorescence multimodality imaging system for small animals via the structured detection FSI architecture. The system is low cost, with a more compact structure, and free of image registration from different modalities. In vivo multimodality imaging results of a mouse bearing tumor demonstrate its capability for small animal research.