Abstract
This chapter describes methods for isolating and imaging metabolically and toxicologically challenged mitochondria with atomic force microscopy. Mitochondria were isolated from rat dorsal root ganglia or brain and exposed to glucose or dinitrobenzene (DNB) to simulate the cellular environment of a diabetic animal that has been exposed to excess glucose or to DNB. It is one of only a few articles to present images of membrane structures, such as voltage-dependent, anion-selective channel pores, on intact organelles. The purpose of the chapter is not to report on the metabolic or toxic effects, but to communicate in more detail than a typical journal paper allows the methods used to image isolated organelles. We also provide a series images revealing the outer membrane and outer membrane pores. An image of an isolated nucleus as well as a set of notes written to avoid common pitfalls in isolation, labeling, and imaging is also included.
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Acknowledgments
The authors thank Michael Bouchard for helpful discussions on mitochondrial structure and function, Chia-Wei Wang and Hui Wang for AFM assistance, Carrie Backus and Terry Miller for mitochondrial isolation assistance, Eva Feldman for funding through the Michigan Diabetes Research and Training Center, Martin Filbert for donation of animal specimens, Ann Marie Sastry for atomic force microscope access, and the Keck Foundation for support.
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Layton, B.E., Boyd, M.B. (2011). Atomic Force Microscopy of Isolated Mitochondria. In: Braga, P., Ricci, D. (eds) Atomic Force Microscopy in Biomedical Research. Methods in Molecular Biology, vol 736. Humana Press. https://doi.org/10.1007/978-1-61779-105-5_10
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DOI: https://doi.org/10.1007/978-1-61779-105-5_10
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