Madl Josef, Rhode Sebastian, Stangl Herbert, Stockinger Hannes, Hinterdorfer Peter, Schütz Gerhard J, Kada Gerald
Institute for Biophysics, Johannes Kepler University Linz, Altenbergerstr. 69, 4040 Linz, Austria.
Ultramicroscopy. 2006 Jun-Jul;106(8-9):645-51. doi: 10.1016/j.ultramic.2005.12.020. Epub 2006 Apr 21.
We present an easy-to-use combination of an atomic force microscope (AFM) and an epi-fluorescence microscope, which allows live cell imaging under physiological conditions. High-resolution AFM images were acquired while simultaneously monitoring either the fluorescence image of labeled membrane components, or a high-contrast optical image (DIC, differential interference contrast). By applying two complementary techniques at the same time, additional information and correlations between structure and function of living organisms were obtained. The synergy effects between fluorescence imaging and AFM were further demonstrated by probing fluorescence-labeled receptor clusters in the cell membrane via force spectroscopy using antibody-functionalized tips. The binding probability on receptor-containing areas identified with fluorescence microscopy ("receptor-positive sites") was significantly higher than that on sites lacking receptors.
我们展示了一种易于使用的原子力显微镜（AFM）和落射荧光显微镜的组合，它能够在生理条件下对活细胞进行成像。在同时监测标记的膜成分的荧光图像或高对比度光学图像（微分干涉差，DIC）的情况下，获取了高分辨率的AFM图像。通过同时应用两种互补技术，获得了关于生物体结构与功能之间的额外信息及相关性。通过使用抗体功能化探针经由力谱法探测细胞膜中荧光标记的受体簇，进一步证明了荧光成像与AFM之间的协同效应。通过荧光显微镜鉴定的含受体区域（“受体阳性位点”）上的结合概率明显高于缺乏受体的位点。
