Light-sheet fluorescence imaging to localize cardiac lineage and protein distribution.
Light-sheet fluorescence imaging to localize cardiac lineage and protein distribution.
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DOI：
10.1038/srep42209
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发表时间：
2017-02-06
影响因子：
4.6
通讯作者：
Hsiai TK
中科院分区：
文献类型：
--
作者：
Ding Y;Lee J;Ma J;Sung K;Yokota T;Singh N;Dooraghi M;Abiri P;Wang Y;Kulkarni RP;Nakano A;Nguyen TP;Fei P;Hsiai TK
光片荧光显微镜 (LSFM) 有助于推进发育研究和再生医学。结合荧光友好组织透明技术的并行进步，我们的心脏 LSFM 能够实现双面照明，从而快速揭示体积超过 10 x 10 x 10mm3 的小鼠心脏的结构；从而可以将祖细胞分化定位到心肌细胞谱系，并以高对比度和分辨率实现 AAV9 介导的外源跨膜钾通道的表达。无需缝合图像列、旋转光片和机械切片心脏的步骤，我们建立了“数字小鼠心脏”的三维重建的整体策略，以评估异常的心脏结构以及新生儿心脏谱系和成人离子通道的空间分布。
Light-sheet fluorescence microscopy (LSFM) serves to advance developmental research and regenerative medicine. Coupled with the paralleled advances in fluorescence-friendly tissue clearing technique, our cardiac LSFM enables dual-sided illumination to rapidly uncover the architecture of murine hearts over 10 by 10 by 10 mm3 in volume; thereby allowing for localizing progenitor differentiation to the cardiomyocyte lineage and AAV9-mediated expression of exogenous transmembrane potassium channels with high contrast and resolution. Without the steps of stitching image columns, pivoting the light-sheet and sectioning the heart mechanically, we establish a holistic strategy for 3-dimentional reconstruction of the “digital murine heart” to assess aberrant cardiac structures as well as the spatial distribution of the cardiac lineages in neonates and ion-channels in adults.