動態三維影像技術於斑馬魚心臟收縮功能定量及鈣離子成像

Dynamic 3D Cardiac Imaging of Zebrafish for the Quantification of Cardiac Function and Calcium Imaging

開發定量模式生物心臟收縮功能及鈣離子成像的技術對各種心臟相關研究，例如：生理學、致病機轉、藥物毒性、發育生物學及再生醫學等，都扮演重要的角色。斑馬魚的胚胎發育快、基因及心臟電生理與人類相近，因此常用於心臟相關研究。斑馬魚幼魚的身體小且透明度高，可直接以光學顯微鏡觀察心臟。但由於其心臟快速跳動，因此獲得精確的心臟收縮功能及鈣離子成像仍是一大挑戰。在此文，我們將介紹如何結合影像處理以及光學成像技術，重建活體斑馬魚心臟腔室的動態三維成像以及動態三維的心臟鈣離子影像。此技術可在同一隻班馬魚同時獲得心室收縮功能參數、跳動心臟中心肌細胞鈣離子瞬態、心臟電生理訊號傳導等重要生理訊息，預期有機會開啟心臟相關全新研究方向及生醫應用。

Quantification of the contractile function and the calcium dynamics of the heart of model organisms is important for research areas ranging from the pathophysiology of heart diseases, the cardiotoxicity of drugs, to the developmental and regenerative biology of the heart. The zebrafish heart develops rapidly and has high genomic and electrophysiological conservation relative to its human counterpart, and hence the zebrafish has long been a popular model animal for cardiac research. While optical imaging of the small and translucent larval zebrafish is straightforward, precise determination of the cardiac function and the calcium dynamic on a rapidly beating heart remains challenging. Here we introduce dynamic three-dimensional (3D) cardiac imaging of larval zebrafish through image reconstruction and demonstrate the simultaneous determination of the cardiac contractile function, the local calcium transient and the electrophysiological conduction on a living zebrafish. In view of the growing interest of using zebrafish in cardiac research, we envisage that our approach should benefit not only contemporary pharmaceutical development but also exploratory research targeting heart diseases.