 Abstract organic small molecule contrast agents have attracted considerable attention in the field of multispectral optoacoustic imaging, but their weak optoacoustic performance resulted from relatively low extinction coefficient and poor water solubility. To overcome this limitation, we constructed supramolecular assemblies based on Q-Kerbit-8, Ural-CB-8, to include two Dixenthean-based chromophores, DXB, and DXB-TZ, which were synthesized as model gas compounds. This led to a substantial enhancement in optoacoustic performance. Furthermore, biological application potential was investigated after co-assembly with chondroitin sulfate A-CSA. The formulated DXB-TZ-CB-8-CSA can effectively detect and diagnose subcutaneous tumours, orthotopic bladder tumours, lymphatic metastasis of tumours, and ischemia, a reperfusion-induced acute kidney injury in mouse models with multispectral optoacoustic imaging. This article was authored by Yinglong Wu, Lehe Sun, Xiaokai Chen, and others. We are article.tv, links in the description below.