Light Sheet Fluorescence Microscopy (LSFM) enables multi-dimensional and multi-scale imaging via illuminating specimens with a separate thin sheet of laser. It allows rapid plane illumination for reduced photo-damage and superior axial resolution and contrast. We hereby demonstrate cardiac LSFM (c-LSFM) imaging to assess the functional architecture of zebrafish embryos with a retrospective cardiac synchronization algorithm for four-dimensional reconstruction (3-D space + time). By combining our approach with tissue clearing techniques, we reveal the entire cardiac structures and hypertrabeculation of adult zebrafish hearts in response to doxorubicin treatment. By integrating the resolution enhancement technique with c-LSFM to increase the resolving power under a large field-of-view, we demonstrate the use of low power objective to resolve the entire architecture of large-scale neonatal mouse hearts, revealing the helical orientation of individual myocardial fibers. Therefore, our c-LSFM imaging approach provides multi-scale visualization of architecture and function to drive cardiovascular research with translational implication in congenital heart diseases.
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机译:薄板荧光显微镜(LSFM)可以通过使用单独的激光薄板照射样品来进行多维和多尺度成像。它可以快速进行平面照明,以减少光损伤,并具有出色的轴向分辨率和对比度。我们在此演示心脏LSFM(c-LSFM)成像,以回顾性心脏同步算法对四维重建(3-D space ++ time)评估斑马鱼胚胎的功能结构。通过将我们的方法与组织清除技术相结合,我们揭示了响应阿霉素治疗的成年斑马鱼心脏的整个心脏结构和超小梁。通过将分辨率增强技术与c-LSFM集成以提高大视野下的分辨能力,我们演示了使用低功率物镜来解决大型新生小鼠心脏的整个结构,揭示了螺旋形的方向单个心肌纤维。因此,我们的c-LSFM成像方法可对结构和功能进行多尺度可视化,以驱动先天性心脏病中具有转化意义的心血管研究。
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