De Stasio Gelsomina, Frazer Bradley H, Girasole Marco, Wiese Lisa M, Krasnowska Ewa K, Greco Giulia, Serafino Annalucia, Parasassi Tiziana
University of Wisconsin-Madison, Department of Physics and Synchrotron Radiation Center, Stoughton, Wisconsin 53589, USA.
Microsc Res Tech. 2004 Feb 1;63(2):115-21. doi: 10.1002/jemt.20019.
Established microscopies such as Scanning Electron Microscopy (SEM) and more recent developments such as Atomic Force Microscopy (AFM) and X-ray Photo-Electron Emission spectroMicroscopy (X-PEEM) can only image the sample surface. We present an argon sputtering method able to progressively expose inner cell structures without apparent damage. By varying the sputtering time, the structure of cell cytoskeleton, vesicles, mitochondria, nuclear membrane, and nucleoli can be imaged. We compared images obtained with confocal fluorescence microscopy, transmission electron microscopy (TEM), SEM, and X-PEEM on similar samples after argon sputtering, then confirmed the similarity of reference intracellular structures, including cytoskeleton fibers, cell-cell and cell-substrate adhesion structures, and secretory vesicles. We conclude that the sputtering method is a new valuable tool for surface sensitive microscopies.
诸如扫描电子显微镜（SEM）等已有的显微镜技术，以及诸如原子力显微镜（AFM）和X射线光电子发射光谱显微镜（X-PEEM）等最新进展，都只能对样品表面进行成像。我们提出了一种氩离子溅射方法，该方法能够逐步揭示细胞内部结构而无明显损伤。通过改变溅射时间，可以对细胞骨架、囊泡、线粒体、核膜和核仁的结构进行成像。我们比较了在氩离子溅射后，用共聚焦荧光显微镜、透射电子显微镜（TEM）、SEM和X-PEEM对相似样品所获得的图像，然后证实了包括细胞骨架纤维、细胞-细胞和细胞-基质粘附结构以及分泌囊泡在内的参考细胞内结构的相似性。我们得出结论，溅射方法是一种对表面敏感显微镜技术而言有价值的新工具。
