Centre d'Imageries Plasmoniques Appliquées, Institut Langevin, Ecole Supérieure de Physique et de Chimie Industrielles (ESPCI) ParisTech, CNRS UMR 7587, Université Paris Diderot, Paris, France.
Phys Rev Lett. 2012 May 25;108(21):218101. doi: 10.1103/PhysRevLett.108.218101. Epub 2012 May 22.
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.
我们报告了一种新的全场荧光显微镜方法,用于基于超临近场发射成像活细胞膜。该技术包括通过简单的图像相减从下临界光中提取超临界光的自干涉。在物镜后焦平面,这等效于添加虚拟掩模来阻挡下临界发射。我们表明,这种虚拟掩模与真实物理掩模截然不同,能够实现 100nm 的轴向约束,并在不损害横向分辨率的情况下提高图像灵敏度。该技术易于实现,同时可以用标准照明光提供细胞内部及其膜的图像。