High-resolution x-ray diffraction microscopy of specifically labeled yeast cells
High-resolution x-ray diffraction microscopy of specifically labeled yeast cells
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DOI：
10.1073/pnas.0910874107
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发表时间：
2010-04-20
影响因子：
11.1
通讯作者：
Jacobsen, Chris
中科院分区：
文献类型：
--
作者：
Nelson, Johanna;Huang, Xiaojing;Jacobsen, Chris
X射线衍射显微镜补充了其他X射线显微镜的方法是免费的镜头强加的辐射剂量和分辨率限制，它允许高分辨率成像的生物标本太厚，以查看电子显微镜。我们在这里报告的最高分辨率（11-13 nm）的X射线衍射显微照片的生物标本，和分子特异性的金标记在不同深度的细胞内通过焦点传播的重建波场的演示。将凝集素伴刀豆球蛋白A与胶体金颗粒结合，用于标记酿酒酵母细胞壁中的α-甘露聚糖糖。将细胞在液体乙烷中速冻并冻干，之后使用X射线衍射显微镜在750 eV光子能量下对它们进行整体成像。
X-ray diffraction microscopy complements other x-ray microscopy methods by being free of lens-imposed radiation dose and resolution limits, and it allows for high-resolution imaging of biological specimens too thick to be viewed by electron microscopy. We report here the highest resolution (11-13 nm) x-ray diffraction micrograph of biological specimens, and a demonstration of molecular-specific gold labeling at different depths within cells via through-focus propagation of the reconstructed wavefield. The lectin concanavalin A conjugated to colloidal gold particles was used to label the a-mannan sugar in the cell wall of the yeast Saccharomyces cerevisiae. Cells were plunge-frozen in liquid ethane and freeze-dried, after which they were imaged whole using x-ray diffraction microscopy at 750 eV photon energy.