Real-time imaging of the surface topography of living yeast cells by atomic force microscopy
Real-time imaging of the surface topography of living yeast cells by atomic force microscopy
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DOI：
10.1002/yea.923
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发表时间：
2003-01-15
期刊：
影响因子：
2.6
通讯作者：
Dufrêne, YF
中科院分区：
文献类型：
--
作者：
Ahimou, FO;Touhami, A;Dufrêne, YF
原子力显微镜（AFM）用于以高分辨率对活的酿酒酵母细胞的表面形貌进行成像，并实时监测细胞壁的酶消化。除了存在芽痕外，在水溶液中成像的天然细胞的表面是均匀且光滑的。记录表面形貌图像的横向分辨率为 2 mn，且表面形态没有明显改变。添加蛋白酶和淀粉葡萄糖苷酶溶液后，以固定的时间间隔收集单细胞的连续图像。蛋白酶导致表面粗糙度逐渐增加。形成了由突出边缘包围的大凹陷，高度类似于50 nm，归因于甘露糖蛋白外层的侵蚀。相比之下，添加淀粉葡萄糖苷酶后没有观察到细胞表面的改变，这与细胞壁生化组成一致。这些结果表明，AFM 是电子显微镜的补充工具，因为它允许直接实时探索活细胞的表面。版权所有 (C) 2002 John Wiley Sons, Ltd.
Atomic force microscopy (AFM) was used to image the surface topography of living Saccharomyces cerevisiae cells at high resolution and to monitor enzyme digestion of the cell wall in real time. Apart from the presence of bud scars, the surface of native cells imaged in aqueous solution was homogeneous and smooth. Topographic images of the surface were recorded to a lateral resolution of 2 mn without significant modification of the surface morphology. Successive images of single cells were collected at fixed time intervals following addition of protease and amyloglucosidase solutions. Protease caused a progressive increase of surface roughness. Large depressions surrounded by protruding edges, similar to50 nm in height, were formed and attributed to the erosion of the mannoprotein outer layer. By contrast, no modification of the cell surface was noted upon addition of amyloglucosidase, which was consistent with the cell wall biochemical composition. These results indicate that AFM is a complementary tool to electron microscopy in that it allows the surface of living cells to be explored directly in real time. Copyright (C) 2002 John Wiley Sons, Ltd.