Advantages of intermediate X-ray energies in Zernike phase contrast X-ray microscopy

被引:9
|
作者
Wang, Zhili [1 ,2 ]
Gao, Kun [1 ]
Chen, Jian [1 ]
Hong, Youli [2 ]
Ge, Xin [1 ]
Wang, Dajiang [1 ]
Pan, Zhiyun [1 ]
Zhu, Peiping [2 ]
Yun, Wenbing [3 ]
Jacobsen, Chris [4 ,5 ]
Wu, Ziyu [1 ,2 ]
机构
[1] Univ Sci & Technol China, Natl Synchrotron Radiat Lab, Hefei 230026, Peoples R China
[2] Chinese Acad Sci, Inst High Energy Phys, Beijing 100049, Peoples R China
[3] Xradia Inc, Pleasanton, CA 94588 USA
[4] Argonne Natl Lab, Adv Photon Source, Argonne, IL 60439 USA
[5] Northwestern Univ, Evanston, IL USA
基金
中国国家自然科学基金; 中国博士后科学基金;
关键词
X-ray microscopy; Zernike phase contrast; Intermediate energy; Tomography; Radiation dose; Depth-of-focus; SCHIZOSACCHAROMYCES-POMBE; BIOLOGICAL SPECIMENS; SPATIAL-RESOLUTION; NM RESOLUTION; TOMOGRAPHY; TRANSMISSION; CELLS; SAMPLES; YEAST; SOIL;
D O I
10.1016/j.biotechadv.2012.04.001
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Understanding the hierarchical organizations of molecules and organelles within the interior of large eukaryotic cells is a challenge of fundamental interest in cell biology. Light microscopy is a powerful tool for observations of the dynamics of live cells, its resolution attainable is limited and insufficient. While electron microscopy can produce images with astonishing resolution and clarity of ultra-thin (<1 mu m thick) sections of biological specimens, many questions involve the three-dimensional organization of a cell or the interconnectivity of cells. X-ray microscopy offers superior imaging resolution compared to light microscopy, and unique capability of nondestructive three-dimensional imaging of hydrated unstained biological cells, complementary to existing light and electron microscopy. Until now, X-ray microscopes operating in the "water window" energy range between carbon and oxygen k-shell absorption edges have produced outstanding 3D images of cryo-preserved cells. The relatively low X-ray energy (<540 eV) of the water window imposes two important limitations: limited penetration (<10 mu m) not suitable for imaging larger cells or tissues, and small depth of focus (DoF) for high resolution 3D imaging (e.g., similar to 1 mu m DoF for 20 nm resolution). An X-ray microscope operating at intermediate energy around 2.5 key using Zernike phase contrast can overcome the above limitations and reduces radiation dose to the specimen. Using a hydrated model cell with an average chemical composition reported in literature, we calculated the image contrast and the radiation dose for absorption and Zernike phase contrast, respectively. The results show that an X-ray microscope operating at similar to 2.5 key using Zernike phase contrast offers substantial advantages in terms of specimen size, radiation dose and depth-of-focus. (C) 2012 Elsevier Inc. All rights reserved.
引用
收藏
页码:387 / 392
页数:6
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