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Immersion autometallographic tracing of zinc ions in Alzheimer beta-amyloid plaques

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Abstract

An easy to perform autometallographic technique (AMG) for capturing zinc ions in Alzheimer plaques is presented. The possibility of visualizing loosely bound or free zinc ions in tissue by immersion autometallography (iZnSAMG) is a relatively recent development. The iZnSAMG staining is caused by zinc-sulphur nanocrystals created in 1–2 mm thick brain slices that are immersed in a 0.1% sodium sulphide, 3% glutaraldehyde phosphate buffered solution, the NeoTimm Solution (NTS), for 3 days. When the zinc-sulphur nanocrystals are subsequently silver-enhanced by autometallography, the plaques are readily identified as spheres of dark interlacing strands of different sizes, embedded in the pattern of zinc-enriched terminals. The zinc specificity of the iZnSAMG technique was tested by immersion of brain slides in the chelator DEDTC prior to the NTS immersion. The iZnSAMG detection of zinc ions is easily standardized and can be used in the quantification of plaques with stereological methods. This technique is the first to detect zinc in plaques in the cerebellum of transgenic PS1/APP mice and the first to detect zinc ions in plaques and dystrophic neurites at electron microscopical levels.

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Acknowledgements

We thank Dr. David R. Borchelt, Department of Pathology, The Johns Hopkins University School of Medicine, for kindly providing the transgenic PS1/APP mice. We also gratefully acknowledge the skillful technical assistance of Ms. D. Jensen, Mr. J. Lund, Mr. A. Meier, Ms. H. Mikkelsen, Dr. O. Pletnikova, Ms. M. Sand, and Ms. K. Wiedemann. This study was supported by The Danish Medical Research Council, the Aarhus University Research Foundation, Aase & Ejnar Danielsens Fond, the Danish Medical Association Research Fund (the Søren Segel & Johanne Wiibroe Segels Research Grant), and the Lundbeck, Leo, Beckett, Gangsted, and Novo Nordic Foundations and by a grant from the National Institute of Health (P50 AG 05146).

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Correspondence to M. Stoltenberg.

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Stoltenberg, M., Bruhn, M., Søndergaard, C. et al. Immersion autometallographic tracing of zinc ions in Alzheimer beta-amyloid plaques. Histochem Cell Biol 123, 605–611 (2005). https://doi.org/10.1007/s00418-005-0787-0

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