Piontek Melissa C, Roos Wouter H
Moleculaire Biofysica, Zernike Instituut, Rijksuniversiteit Groningen, Nijenborgh 4, 9747 AG, Groningen, The Netherlands.
Methods Mol Biol. 2018;1665:243-258. doi: 10.1007/978-1-4939-7271-5_13.
Imaging of nano-sized particles and sample features is crucial in a variety of research fields. For instance in biological sciences, where it is paramount to investigate structures at the single particle level. Often two-dimensional images are not sufficient and further information such as topography and mechanical properties are required. Furthermore, to increase the biological relevance, it is desired to perform the imaging in close to physiological environments. Atomic force microscopy (AFM) meets these demands in an all-in-one instrument. It provides high-resolution images including surface height information leading to three-dimensional information on sample morphology. AFM can be operated both in air and in buffer solutions. Moreover, it has the capacity to determine protein and membrane material properties via the force spectroscopy mode. Here we discuss the principles of AFM operation and provide examples of how biomolecules can be studied. By including new approaches such as high-speed AFM (HS-AFM) we show how AFM can be used to study a variety of static and dynamic single biomolecules and biomolecular assemblies.
纳米级颗粒和样品特征的成像在各种研究领域中至关重要。例如在生物科学领域，在单颗粒水平上研究结构至关重要。通常二维图像并不足够，还需要诸如形貌和机械性能等进一步信息。此外，为了提高生物学相关性，希望在接近生理环境中进行成像。原子力显微镜（AFM）在一台仪器中满足了这些需求。它提供高分辨率图像，包括表面高度信息，从而得到关于样品形态的三维信息。AFM 既可以在空气中操作，也可以在缓冲溶液中操作。此外，它有能力通过力谱模式确定蛋白质和膜材料的性质。在这里，我们讨论 AFM 的操作原理，并提供如何研究生物分子的示例。通过纳入诸如高速 AFM（HS-AFM）等新方法，我们展示了 AFM 如何用于研究各种静态和动态的单个生物分子以及生物分子组装体。
