细胞骨架元素的原子力显微镜可视化
Visualization of Cytoskeletal Elements by the Atomic Force Microscope
摘要：
我们描述了一种新的应用原子力显微镜（AFM）直接可视化细胞骨架纤维在人类包皮上皮细胞。使用低浓度的非离子去污剂Triton X-100从细胞中去除膜、可溶性蛋白和细胞器。然后，剩余的细胞骨架可以在液体或空气干燥的环境条件下直接可视化。这两种类型的扫描提供了补充信息。在液体中扫描可显示细胞骨架的表面细丝，而在空气中扫描可显示细胞骨架纤维的表面细丝和总“体积”。观察到的最小纤维为ca。直径为50 nm。该技术的横向分辨率为约。20 nm，这可以增加到一个单一的纳米级，通过选择更尖锐的AFM针尖。由于AFM是一种真正的三维技术，我们能够通过其密度和体积来量化所观察到的细胞骨架。纤维的类型可以通过它们的大小来识别，类似于电子显微镜。
英文摘要：
We describe a novel application of atomic force microscopy (AFM) to directly visualize cytoskeletal fibers in human foreskin epithelial cells. The nonionic detergent Triton X-100 in a low concentration was used to remove the membrane, soluble proteins, and organelles from the cell. The remaining cytoskeleton can then be directly visualized in either liquid or air-dried ambient conditions. These two types of scanning provide complimentary information. Scanning in liquid visualizes the surface filaments of the cytoskeleton, whereas scanning in air shows both the surface filaments and the total "volume" of the cytoskeletal fibers. The smallest fibers observed were ca. 50 nm in diameter. The lateral resolution of this technique was ca.20 nm, which can be increased to a single nanometer level by choosing sharper AFM tips. Because the AFM is a true three dimensional technique, we are able to quantify the observed cytoskeleton by its density and volume. The types of fibers can be identified by their size, similar to electron microscopy.
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