Höppener Christiane, Novotny Lukas
The Institute of Optics and Department of Biomedical Engineering, University of Rochester, Rochester, NY 14627, USA.
Nanotechnology. 2008 Sep 24;19(38):384012. doi: 10.1088/0957-4484/19/38/384012. Epub 2008 Aug 12.
The localization and identification of individual proteins is of key importance for the understanding of biological processes on the molecular scale. Here, we demonstrate near-field fluorescence imaging of single proteins in their native cell membrane. Incident laser radiation is localized and enhanced with an optical antenna in the form of a spherical gold particle attached to a pointed dielectric tip. Individual proteins can be identified with a diffraction-unlimited spatial resolution of ∼50 nm. Besides determining the concentration and distribution of specific membrane proteins, this approach makes it possible to study the colocalization of different membrane proteins. Moreover, it enables a simultaneous recording of the membrane topology. Protein distributions can be correlated with the local membrane topology, thereby providing important information on the chemical and structural organization of cellular membranes.
单个蛋白质的定位和识别对于在分子尺度上理解生物过程至关重要。在此,我们展示了天然细胞膜中单个蛋白质的近场荧光成像。入射激光辐射通过附着在尖锐介电尖端上的球形金颗粒形式的光学天线进行定位和增强。单个蛋白质可以用约50纳米的衍射极限空间分辨率进行识别。除了确定特定膜蛋白的浓度和分布外,这种方法还能够研究不同膜蛋白的共定位。此外,它还能同时记录膜拓扑结构。蛋白质分布可以与局部膜拓扑结构相关联,从而提供有关细胞膜化学和结构组织的重要信息。