I5S:: Wide-field light microscopy with 100-nm-scale resolution in three dimensions
I5S:: Wide-field light microscopy with 100-nm-scale resolution in three dimensions
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DOI：
10.1529/biophysj.107.120352
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发表时间：
2008-06-15
影响因子：
3.4
通讯作者：
Gustafsson, Mats G. L.
中科院分区：
文献类型：
--
作者：
Shao, Lin;Isaac, Berith;Gustafsson, Mats G. L.
描述了一种新型的广域荧光显微镜，它通过在具有两个相对物镜的显微镜中使用结构照明来在所有三个维度上产生100纳米尺度的空间分辨率。照明光被一个光栅和一个分束器分成六个相互相干的光束，其中三个通过每个物镜进入样品。所得到的照明强度图案包含轴向和横向的高空间频率分量。此外，发射由两个物镜相干地收集，并在单个相机上以干涉测量方式组合，从而产生具有轴向延伸支撑的检测传递函数。这两种效应结合在一起，产生了近乎各向同性的分辨率。测试样品和生物样品的实验图像证实了理论预测。
Anew type of wide-field fluorescence microscopy is described, which produces 100-nm-scale spatial resolution in all three dimensions, by using structured illumination in a microscope that has two opposing objective lenses. Illumination light is split by a grating and a beam splitter into six mutually coherent beams, three of which enter the specimen through each objective lens. The resulting illumination intensity pattern contains high spatial frequency components both axially and laterally. In addition, the emission is collected by both objective lenses coherently, and combined interferometrically on a single camera, resulting in a detection transfer function with axially extended support. These two effects combine to produce near-isotropic resolution. Experimental images of test samples and biological specimens confirm the theoretical predictions.