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HFE organs become overloaded with iron which causes damage. Iron-overload is commonly detected by NMR imaging, but the spectroscopic technique is insensitive to diamagnetic iron. Here, we used M\u00f6ssbauer spectroscopy to examine the iron content of liver, spleen, kidney, heart, and brain of <jats:sup>57<\/jats:sup>Fe-enriched HFE<jats:sup>(\u2212\/\u2212)<\/jats:sup> mice of ages 3\u201352 wk. Overall, the iron contents of all investigated HFE organs were similar to the same healthy organ <jats:italic>but from an older mouse<\/jats:italic>. Livers and spleens were majorly overloaded, followed by kidneys. Excess iron was generally present as ferritin. Iron\u2013sulfur clusters and low-spin Fe<jats:sup>II<\/jats:sup> hemes (combined into the central quadrupole doublet) and nonheme high-spin Fe<jats:sup>II<\/jats:sup> species were also observed. Spectra of young and middle-aged HFE kidneys were dominated by the central quadrupole doublet and were largely devoid of ferritin. Collecting and comparing\u00a0spectra at 5 and 60\u00a0K allowed the presence of hemosiderin, a decomposition product of ferritin, to be quantified, and it also allowed the diamagnetic central doublet to be distinguished from ferritin. Hemosiderin was observed in spleens and livers from HFE mice, and in spleens from controls, but only when iron concentrations exceeded 2\u20133\u00a0mM. Even in those cases, hemosiderin represented only 10\u201320% of the iron in the sample. NMR imaging can identify iron-overload under non-invasive room-temperature conditions, but M\u00f6ssbauer spectroscopy of <jats:sup>57<\/jats:sup>Fe-enriched mice can detect all forms of iron and perhaps allow the process of iron-overloading to be probed in greater detail.<\/jats:p>\n                <jats:p><jats:bold>Graphical Abstract<\/jats:bold><\/jats:p>","DOI":"10.1007\/s00775-022-01975-y","type":"journal-article","created":{"date-parts":[[2022,12,13]],"date-time":"2022-12-13T08:03:20Z","timestamp":1670918600000},"page":"173-185","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Low-temperature M\u00f6ssbauer spectroscopy of organs from 57Fe-enriched HFE(\u2212\/\u2212) hemochromatosis mice: an\u00a0iron-dependent threshold for generating hemosiderin"],"prefix":"10.1007","volume":"28","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5091-7405","authenticated-orcid":false,"given":"Shaik Waseem","family":"Vali","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8307-9647","authenticated-orcid":false,"given":"Paul A.","family":"Lindahl","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2022,12,13]]},"reference":[{"issue":"8","key":"1975_CR1","doi-asserted-by":"publisher","first-page":"731","DOI":"10.21037\/atm-20-5512","volume":"9","author":"GJ Anderson","year":"2021","unstructured":"Anderson GJ, Bardou-Jacquet E (2021) Revisiting hemochromatosis: genetic vs. phenotypic manifestations. 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