Three-dimensional integrated quantitative modeling and fluorescent imaging of doxorubicin-induced cardiotoxicity in a
1Key Laboratory of Biotechnology of Antibiotics, the National Health Commission (NHC), Bejing Key Laboratory of Antimicrobial Agents, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Tian Tan Xi Li No.1, Dong Cheng District, Beijing 100050, China.
Abstract:
The quantification and visualization of fluorescent staining at the whole organ level remain a challenge. Deconvolution image systems allow multi-dimensional imaging and stereo-measurement via rapid 3D reconstruction. To demonstrate this technique, we investigated doxorubicin-induced cardiotoxicity in zebrafish. Fluorogenic probe and immunofluorescence were employed to identify cardiac reactive oxygen species generation and myocardial apoptosis, respectively. We revealed the spatial distribution of fluorescent staining across the whole heart by this approach. In addition, the fluorescence intensities and fluorescence-dyed volumes in the zebrafish heart were quantified automatically. Importantly, doxorubicin treatment induced more ROS generation in the ventricle as compared to the atrium, while the levels of activated caspase-3 were much higher in the atrioventricular junction. These results would have been difficult to observe using traditional 2D images. Therefore, our deconvolution imaging strategy allows the 3D quantification and visualization of fluorescent staining at the whole organ level, and will thus support in vivo studies.
