Abstract
This protocol utilizes immunohistochemistry to assist the laser microdissection and capture of tissue regions of interest isolated from slide-mounted sections for subsequent analysis of metal content. When used in conjunction with inductively coupled plasma mass spectrometry it becomes a powerful method to determine pathogenetically related shifts in the concentration of various cortical transition metals. Key advantages of this approach include being able to isolate target and directly adjacent brain regions for easy comparative analysis free from potential metal contamination as can occur during surgically based isolation of tissue elements. In this chapter, I present a well-optimized and validated method to achieve contamination-free detection of microscopically defined regions of manganese concentration in Alzheimer’s disease brain tissue with an emphasis on amyloid plaque analysis.
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Acknowledgments
Confirmed Alzheimer’s and age-matched healthy control brain tissue blocks were obtained through the Victorian Brain Bank Network, Victoria, Australia; Anti-Aβ polyclonal hybridoma supernatant was a kind gift of Dr. QX Li, Department of Pathology, University of Melbourne; ICP-MS analyses were conducted by Ms. Irene Volitakis, MHRI, University of Melbourne; Laser microscopy was performed using Biological Optical Microscopy Platform, University of Melbourne Zeiss equipment. The image in Fig. 2 was generously provided by Carl Zeiss Microscopy GmbH; Manuscript editing assistance was provided by Everyedit, www.everyedit.com
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Brazier, M.W. (2017). Microdissection of Alzheimer Brain Tissue for the Determination of Focal Manganese Accumulation. In: White, A. (eds) Metals in the Brain. Neuromethods, vol 124. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6918-0_6
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DOI: https://doi.org/10.1007/978-1-4939-6918-0_6
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