Shibata Mikihiro, Uchihashi Takayuki, Ando Toshio, Yasuda Ryohei
1] Max Planck Florida Institute for Neuroscience, Jupiter, FL 33458, USA [2] Department of Neurobiology, Duke University Medical School, Durham, NC 27710, USA.
1] Department of Physics, Kanazawa University, Kanazawa 920-1192, Japan [2] Bio-AFM Frontier Research Center, Kanazawa University, Kanazawa 920-1192, Japan [3] CREST/JST, Tokyo 102-0075, Japan.
Sci Rep. 2015 Mar 4;5:8724. doi: 10.1038/srep08724.
Visualization of morphological dynamics of live cells with nanometer resolution under physiological conditions is highly desired, but challenging. It has been demonstrated that high-speed atomic force microscopy is a powerful technique for visualizing dynamics of biomolecules under physiological conditions. However, application of high-speed atomic force microscopy for imaging larger objects such as live mammalian cells has been complicated because of the collision between the cantilever and samples. Here, we demonstrate that attaching an extremely long (3 μm) and thin (5 nm) tip by amorphous carbon to the cantilever allows us to image the surface structure of live cells with the spatiotemporal resolution of nanometers and seconds. We demonstrate that long-tip high-speed atomic force microscopy is capable of imaging morphogenesis of filopodia, membrane ruffles, pit formation, and endocytosis in COS-7, HeLa cells and hippocampal neurons.
在生理条件下以纳米分辨率可视化活细胞的形态动力学是非常需要的，但具有挑战性。已经证明，高速原子力显微镜是一种在生理条件下可视化生物分子动力学的强大技术。然而，由于悬臂与样品之间的碰撞，高速原子力显微镜在对诸如活哺乳动物细胞等较大物体进行成像时变得复杂。在这里，我们证明通过非晶碳将极长（约3μm）且极薄（约5nm）的尖端连接到悬臂上，使我们能够以纳米和秒的时空分辨率对活细胞的表面结构进行成像。我们证明长尖端高速原子力显微镜能够对COS-7细胞、HeLa细胞和海马神经元中的丝状伪足形态发生、膜皱褶、凹陷形成和内吞作用进行成像。
