U1006 INSERM, Université Aix-Marseille, Parc Scientifique de Luminy, Marseille, F-13009, France.
J Mol Recognit. 2012 May;25(5):292-8. doi: 10.1002/jmr.2187.
Atomic force microscopy (AFM) image acquisition is performed by raster-scanning a faint tip with respect to the sample by the use of a piezoelectric stage that is guided by a feedback system. This process implies that the resulting images feature particularities that distinguish them from images acquired by other techniques, such as the drift of the piezoelectric elements, the unequal image contrast along the fast- and the slow-scan axes, the physical contact between the tip of nondefinable geometry and the sample, and the feedback parameters. Recently, high-speed AFM (HS-AFM) has been introduced, which allows image acquisition about three orders of magnitude faster (500-100 ms frame rate) than conventional AFM (500 s to 100 s frame rate). HS-AFM produces image sequences, large data sets, which report biological sample dynamics. To analyze these movies, we have developed a software package that (i) adjusts individual scan lines and images to a common contrast and z-scale, (ii) filters specifically those scan lines where increased or insufficient force was applied, (iii) corrects for piezo-scanner drift, (iv) defines particle localization and angular orientation, and (v) performs particle tracking to analyze the lateral and rotation displacement of single molecules.
原子力显微镜（AFM）图像采集是通过使用压电台在反馈系统的引导下相对于样品进行微探针的光栅扫描来完成的。这一过程意味着所得到的图像具有一些特殊之处，使其与其他技术获得的图像区分开来，例如压电元件的漂移、快扫和慢扫轴上图像对比度的不均匀性、非定形几何形状的探针与样品之间的物理接触以及反馈参数。最近，引入了高速原子力显微镜（HS-AFM），它允许图像采集速度比传统原子力显微镜快三个数量级（500-100ms 帧速率）。HS-AFM 产生图像序列，大数据集，报告生物样本动态。为了分析这些电影，我们开发了一个软件包，该软件包（i）将个别扫描线和图像调整到共同的对比度和 z 比例尺，（ii）专门过滤施加了增加或不足的力的扫描线，（iii）校正压电机架漂移，（iv）定义粒子定位和角取向，（v）进行粒子跟踪以分析单个分子的横向和旋转位移。
