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Journal of Electron Microscopy Advance Access published online on June 14, 2006
Journal of Electron Microscopy, doi:10.1093/jmicro/dfl013
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received February 15, 2006
Accepted April 24, 2006

Full-length: Biological

Three-dimensional structural changes in living hippocampal neurons imaged using magnetic AC mode atomic force microscopy

Sun Yunxu 1, Lin Danying 1, Rui Yanfang 2, Han Dong 3, and Ma Wanyun 1 *

1 The Key Laboratory of Atomic and Molecular Nanosciences of Education Ministry, Department of Physics, Tsinghua University, Zhongguancun, Beijing 100084, China
2 Department of Biological Science and Biotechnology, State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University, Beijing 100084, China
3 The Key Laboratory of Atomic and Molecular Nanosciences of Education Ministry, Department of Physics, Tsinghua University, Zhongguancun, Beijing 100084, China; National Center for Nanoscience and Technology, Beijing 100080, China

* To whom correspondence should be addressed.
Ma Wanyun, E-mail: mawy{at}lsad.tsinghua.edu.cn


  Abstract

We developed the magnetic AC (MAC) mode atomic force microscopy (AFM) to image the 3D ultrastructure of living hippocampal neurons under physiological conditions. Initially, the soma, the dendrites and the growth cones of hippocampal neurons were imaged. The imaging force was adjusted to a small value for the long-term observation. The neural spines were damaged when the tip produced a large force; the spines regenerated after the force was reduced. Subsequently, we explored the relationship between structural changes in hippocampal neurons and Alzheimer's disease by employing the new imaging technique. Time-lapse image acquisition (10 min intervals) showed that the growth cone collapsed after the addition amyloid peptide fragment {beta}(25-35), which is thought to initiate Alzheimer's disease. In addition, we found substantial changes in mechanical properties and in the volume of individual growth cone. This study suggested that MAC mode AFM may be a powerful tool for observing long-term structural changes in living neural cells under physiological conditions.

Keywords: hippocampal neuron; regeneration; growth cone; atomic force microscopy.
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