Full-Field Near-Field Optical Microscope for Cell Imaging
Full-Field Near-Field Optical Microscope for Cell Imaging
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DOI：
10.1103/physrevlett.108.218101
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发表时间：
2012-05-22
影响因子：
8.6
通讯作者：
Fort, Emmanuel
中科院分区：
文献类型：
--
作者：
Barroca, Thomas;Balaa, Karla;Fort, Emmanuel
我们报道了一种新的基于超临界近场发射的活细胞膜全场荧光显微成像方法。该技术通过简单的图像相减提取亚临界和超临界光之间的自干涉。在物镜后焦平面中，这相当于添加了阻挡亚临界发射的虚拟掩模。我们表明，这个虚拟的掩模是从根本上不同于一个真实的物理掩模，使100 nm的轴向限制和提高图像灵敏度，而不损害横向分辨率。该技术易于实现，并同时提供细胞内部及其膜的图像与标准照明光。
We report a new full-field fluorescence microscopy method for imaging live cell membranes based on supercritical near-field emission. This technique consists of extracting the self-interference between undercritical and supercritical light by simple image subtraction. In the objective back focal plane, this is equivalent to adding a virtual mask blocking the subcritical emission. We show that this virtual mask is radically different from a real physical mask, enabling a 100 nm axial confinement and enhancing the image sensitivity without damaging the lateral resolution. This technique is easy to implement and simultaneously provides images of the inner parts of the cell and its membrane with standard-illumination light.