A novel optical microscope for imaging large embryos and tissue volumes with sub-cellular resolution throughout
A novel optical microscope for imaging large embryos and tissue volumes with sub-cellular resolution throughout
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DOI：
10.7554/elife.18659
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发表时间：
2016-09-23
期刊：
影响因子：
7.7
通讯作者：
Amos, William Bradshaw
中科院分区：
文献类型：
--
作者：
McConnell, Gail;Tragardh, Johanna;Amos, William Bradshaw
目前的光学显微镜物镜的低放大倍率具有低的数值孔径，因此具有太小的深度分辨率和歧视，以及在共焦和非线性显微镜。这在重要领域是一个严重的限制，包括通过研究经历高级器官形成的胚胎对转基因小鼠中的人类基因进行表型筛选。我们已经建立了一个光学透镜系统，用于对宽达6 mm、厚达3 mm的物体进行3D成像，其深度分辨率仅为几微米，而不是目前达到的几十微米，从而可以在整个体积中解析亚细胞细节。我们提出这个透镜，称为Mesolens，与性能数据和图像从生物标本，包括整个固定和完整的荧光染色的12.5天的小鼠胚胎的共聚焦图像。
Current optical microscope objectives of low magnification have low numerical aperture and therefore have too little depth resolution and discrimination to perform well in confocal and nonlinear microscopy. This is a serious limitation in important areas, including the phenotypic screening of human genes in transgenic mice by study of embryos undergoing advanced organogenesis. We have built an optical lens system for 3D imaging of objects up to 6 mm wide and 3 mm thick with depth resolution of only a few microns instead of the tens of microns currently attained, allowing sub-cellular detail to be resolved throughout the volume. We present this lens, called the Mesolens, with performance data and images from biological specimens including confocal images of whole fixed and intact fluorescently-stained 12.5-day old mouse embryos.