3D visualization of subcellular structures of Schizosaccharomyces pombe by hard X-ray tomography.
Cellular structures of the fission yeast, Schizosaccharomyces pombe, were examined by using hard X-ray tomography. Since cells are nearly transparent to hard X-rays, Zernike phase contrast and heavy metal staining were introduced to improve image contrast. Through using such methods, images taken at 8 keV displayed sufficient contrast for observing cellular structures. The cell wall, the intracellular organelles and the entire structural organization of the whole cells were visualized in three-dimensional at a resolution better than 100 nm. Comparison between phase contrast and absorption contrast was also made, indicating the obvious advantage of phase contrast for cellular imaging at this energy. Our results demonstrate that hard X-ray tomography with Zernike phase contrast is suitable for cellular imaging. Its unique abilities make it have potential to become a useful tool for revealing structural information from cells, especially thick eukaryotic cells.
National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, Anhui, People's Republic of China.
Y
Y
Yang
W
W
Li
G
G
Liu
X
X
Zhang
J
J
Chen
W
W
Wu
Y
Y
Guan
Y
Y
Xiong
Y
Y
Tian
Z
Z
Wu
D002473:Cell Wall / Q000648:ultrastructure; D003952:Diagnostic Imaging / Q000379:methods; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000295:instrumentation* / Q000379:methods*; D019216:Metals, Heavy; D008858:Microscopy, Phase-Contrast; D015388:Organelles / Q000648:ultrastructure; D012568:Schizosaccharomyces / Q000648:ultrastructure*; D013194:Staining and Labeling / Q000379:methods; D014057:Tomography, X-Ray Computed / Q000379:methods*
D019216:Metals, Heavy