Institute of Biophysics, Johannes Kepler University, A-4020 Linz, Austria.
Micron. 2012 Dec;43(12):1399-407. doi: 10.1016/j.micron.2012.05.007. Epub 2012 Jun 3.
In this study, we demonstrate the increased performance in speed and sensitivity achieved by the use of small AFM cantilevers on a standard AFM system. For this, small rectangular silicon oxynitride cantilevers were utilized to arrive at faster atomic force microscopy (AFM) imaging times and more sensitive molecular recognition force spectroscopy (MRFS) experiments. The cantilevers we used had lengths between 13 and 46 μm, a width of about 11 μm, and a thickness between 150 and 600 nm. They were coated with chromium and gold on the backside for a better laser reflection. We characterized these small cantilevers through their frequency spectrum and with electron microscopy. Due to their small size and high resonance frequency we were able to increase the imaging speed by a factor of 10 without any loss in resolution for images from several μm scansize down to the nanometer scale. This was shown on bacterial surface layers (s-layer) with tapping mode under aqueous, near physiological conditions and on nuclear membranes in contact mode in ambient environment. In addition, we showed that single molecular forces can be measured with an up to 5 times higher force sensitivity in comparison to conventional cantilevers with similar spring constants.
在这项研究中,我们展示了在标准原子力显微镜系统上使用小型原子力显微镜悬臂梁可以提高速度和灵敏度。为此,我们使用了小矩形氮化硅悬臂梁,以实现更快的原子力显微镜(AFM)成像时间和更灵敏的分子识别力谱(MRFS)实验。我们使用的悬臂梁长度在 13 到 46 μm 之间,宽度约为 11 μm,厚度在 150 到 600 nm 之间。它们的背面涂有铬和金,以提高激光反射率。我们通过它们的频谱和电子显微镜来表征这些小悬臂梁。由于它们的尺寸小且共振频率高,我们能够将成像速度提高 10 倍,而不会对几 μm 扫描尺寸的图像分辨率造成任何损失,甚至可以达到纳米级。这在水相、接近生理条件下的细菌表面层(S-层)和环境中的接触模式核膜的敲击模式下得到了证明。此外,我们还表明,与具有类似弹性常数的传统悬臂梁相比,我们可以以高达 5 倍的力灵敏度测量单分子力。