3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy.
3D imaging of the early embryonic chicken heart with focused ion beam scanning electron microscopy.
复制标题
使用聚焦离子束扫描电子显微镜对早期胚胎鸡心脏进行 3D 成像。
DOI：
10.1017/s1431927614000828
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Rugonyi,Sandra
中科院分区：
文献类型：
--
作者：
Rennie,MoniqueY;Gahan,CurranG;López,ClaudiaS;Thornburg,KentL;Rugonyi,Sandra
Early embryonic heart development is a period of dynamic growth and remodeling, with rapid changes occurring at the tissue, cell, and subcellular levels. A detailed understanding of the events that establish the components of the heart wall has been hampered by a lack of methodologies for three-dimensional (3D), high-resolution imaging. Focused ion beam scanning electron microscopy (FIB-SEM) is a novel technology for imaging 3D tissue volumes at the subcellular level. FIB-SEM alternates between imaging the block face with a scanning electron beam and milling away thin sections of tissue with a FIB, allowing for collection and analysis of 3D data. FIB-SEM was used to image the three layers of the day 4 chicken embryo heart: myocardium, cardiac jelly, and endocardium. Individual images obtained with FIB-SEM were comparable in quality and resolution to those obtained with transmission electron microscopy. Up to 1,100 serial images were obtained in 4 nm increments at 4.88 nm resolution, and image stacks were aligned to create volumes 800–1,500μm3in size. Segmentation of organelles revealed their organization and distinct volume fractions between cardiac wall layers. We conclude that FIB-SEM is a powerful modality for 3D subcellular imaging of the embryonic heart wall.
