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The effect of formalin fixation on the levels of brain transition metals in archived samples

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Abstract

Reports that iron, zinc and copper homeostasis are in aberrant homeostasis are common for various neurodegenerative diseases, particularly for Huntington’s disease, Parkinson’s disease, and Alzheimer’s disease. Manipulating the levels of these elements in the brain through the application of chelators has been and continues to be tested therapeutically in clinical trials with mixed results. Much of the data indicating that these metals are abnormally concentrated in Alzheimer’s disease and Parkinson’s disease brain tissue was generated through the analysis of post-mortem human tissue which was archived in formalin. In this study, we evaluated the effect of formalin fixation of brain on the levels of three important transition metals (iron, copper, and zinc) by atomic absorption spectroscopy. Paired brain specimens were obtained at autopsy for each case; one was conserved by formalin archival (averaging four years), the other was rapidly frozen. Both white and grey matter samples were analyzed and the concentrations of iron and zinc were found to decrease with fixation. Iron was reduced by 40% (P < 0.01), and zinc by 77% (P < 0.0001); copper concentrations increased by 37% (P < 0.05) by the paired T-test. The increase in copper is likely due to contamination from trace copper in the formalin. These results indicate that transition metal data obtained from fixed tissue may be heavily distorted and care should be taken in interpreting this data.

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Acknowlegements

This research was funded by the National Institutes of Health (AG20948). Harry V. Vinters is supported in part by P01 AG12435, P50 AG16570 and the Daljit S. and Elaine Sarkaria Chair in Diagnostic Medicine. None of the authors have real or potential conflicts of interest related to this work.

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Authors and Affiliations

  1. Neurosurgery Center for Research, Loma Linda University, 11175 Campus Street, Coleman Pavilion Suite 11113, Loma Linda, CA, 92350, USA

    Matthew Schrag, April Dickson, Arshad Jiffry, David Kirsch & Wolff Kirsch

  2. Department of Pathology and Laboratory Medicine, University of California Los Angeles, Los Angeles, CA, USA

    Harry V. Vinters

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  1. Matthew Schrag
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  2. April Dickson
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  3. Arshad Jiffry
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  4. David Kirsch
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  5. Harry V. Vinters
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  6. Wolff Kirsch
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Correspondence to Matthew Schrag.

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Schrag, M., Dickson, A., Jiffry, A. et al. The effect of formalin fixation on the levels of brain transition metals in archived samples. Biometals 23, 1123–1127 (2010). https://doi.org/10.1007/s10534-010-9359-4

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  • DOI: https://doi.org/10.1007/s10534-010-9359-4

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