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Nanoscale X-Ray Microscopic Imaging of Mammalian Mineralized Tissue

Published online by Cambridge University Press:  07 April 2010

Joy C. Andrews*
Affiliation:
Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA
Eduardo Almeida
Affiliation:
NASA Ames Research Center, Moffett Field, CA 94035, USA
Marjolein C.H. van der Meulen
Affiliation:
Sibley School of Mechanical and Aerospace Engineering, Cornell University, Ithaca, NY 14853, USA
Joshua S. Alwood
Affiliation:
Department of Aeronautics and Astronautics, Stanford University, Stanford, CA 94305, USA
Chialing Lee
Affiliation:
Department of Biological Science, San Jose State University, San Jose, CA 95192, USA
Yijin Liu
Affiliation:
Institute of High Energy Physics, Beijing, China
Jie Chen
Affiliation:
National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, China
Florian Meirer
Affiliation:
Institute for Atomic and Subatomic Physics, Technical University of Vienna, Austria
Michael Feser
Affiliation:
Xradia Inc., Concord, CA 94520, USA
Jeff Gelb
Affiliation:
Xradia Inc., Concord, CA 94520, USA
Juana Rudati
Affiliation:
Xradia Inc., Concord, CA 94520, USA
Andrei Tkachuk
Affiliation:
Xradia Inc., Concord, CA 94520, USA
Wenbing Yun
Affiliation:
Xradia Inc., Concord, CA 94520, USA
Piero Pianetta
Affiliation:
Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA
*
Corresponding author. E-mail: jandrews@slac.stanford.edu
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Abstract

A novel hard transmission X-ray microscope (TXM) at the Stanford Synchrotron Radiation Lightsource operating from 5 to 15 keV X-ray energy with 14 to 30 μm2 field of view has been used for high-resolution (30–40 nm) imaging and density quantification of mineralized tissue. TXM is uniquely suited for imaging of internal cellular structures and networks in mammalian mineralized tissues using relatively thick (50 μm), untreated samples that preserve tissue micro- and nanostructure. To test this method we performed Zernike phase contrast and absorption contrast imaging of mouse cancellous bone prepared under different conditions of in vivo loading, fixation, and contrast agents. In addition, the three-dimensional structure was examined using tomography. Individual osteocytic lacunae were observed embedded within trabeculae in cancellous bone. Extensive canalicular networks were evident and included processes with diameters near the 30–40 nm instrument resolution that have not been reported previously. Trabecular density was quantified relative to rod-like crystalline apatite, and rod-like trabecular struts were found to have 51–54% of pure crystal density and plate-like areas had 44–53% of crystal density. The nanometer resolution of TXM enables future studies for visualization and quantification of ultrastructural changes in bone tissue resulting from osteoporosis, dental disease, and other pathologies.

Type
Biological Applications
Copyright
Copyright © Microscopy Society of America 2010

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References

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