title: Biochemical and biophysical methods for studying mitochondrial iron metabolism.
abstract: Iron is a heavily utilized element in organisms and numerous mechanisms accordingly regulate the trafficking, metabolism, and storage of iron. Despite the high regulation of iron homeostasis, several diseases and mutations can lead to the misregulation and often accumulation of iron in the cytosol or mitochondria of tissues. To understand the genesis of iron overload, it is necessary to employ various techniques to quantify iron in organisms and mitochondria. This chapter discusses techniques for determining the total iron content of tissue samples, ranging from colorimetric determination of iron concentrations, atomic absorption spectroscopy, inductively coupled plasma-optical emission spectroscopy, and inductively coupled plasma-mass spectrometry. In addition, we discuss in situ techniques for analyzing iron including electron microscopic nonheme iron histochemistry, electron energy loss spectroscopy, synchrotron X-ray fluorescence imaging, and confocal Raman microscopy. Finally, we discuss biophysical methods for studying iron in isolated mitochondria, including ultraviolet-visible, electron paramagnetic resonance, X-ray absorbance, and Mössbauer spectroscopies. This chapter should aid researchers to select and interpret mitochondrial iron quantifications.
authors.affiliation: Molecular Medicine Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, Maryland, USA.
authors.forename: Gregory P
authors.initials: GP
authors.lastname: Holmes-Hampton
authors.affiliation: Molecular Medicine Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, Maryland, USA.
authors.forename: Wing-Hang
authors.initials: WH
authors.lastname: Tong
authors.affiliation: Molecular Medicine Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, Maryland, USA. Electronic address: rouault@mail.nih.gov.
authors.forename: Tracey A
authors.initials: TA
authors.lastname: Rouault
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods*; D001703:Biophysics / Q000379:methods*; D003124:Colorimetry / Q000379:methods; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D006801:Humans; D007501:Iron / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D008928:Mitochondria / Q000378:metabolism*; D013054:Spectrophotometry, Atomic / Q000379:methods; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D015204:Spectroscopy, Mossbauer / Q000379:methods; D013057:Spectrum Analysis / Q000379:methods; D013059:Spectrum Analysis, Raman / Q000379:methods
chemical_list: D007501:Iron
keywords: AAS; EELS; EPR spectroscopy; ICP-MS; ICP-OES; Iron metabolism; Mössbauer spectroscopy; Perls’ DAB; Perls’ stain; UV-vis spectroscopy; XAS