{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T12:39:10Z","timestamp":1791290350487,"version":"4.3.0"},"reference-count":31,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2023,12,26]],"date-time":"2023-12-26T00:00:00Z","timestamp":1703548800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100004704","name":"National Research Council of Thailand","doi-asserted-by":"publisher","award":["N41A650085"],"award-info":[{"award-number":["N41A650085"]}],"id":[{"id":"10.13039\/501100004704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006251","name":"Universit\u00e9 de Bordeaux","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100006251","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,1,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Iron dyshomeostasis is involved in many neurological disorders, particularly neurodegenerative diseases where iron accumulates in various brain regions. Identifying mechanisms of iron transport in the brain is crucial for understanding the role of iron in healthy and pathological states. In neurons, it has been suggested that iron can be transported by the axon to different brain regions in the form of labile iron; a pool of reactive and exchangeable intracellular iron. Here we report a novel approach to imaging labile ferrous iron, Fe(II), in live primary hippocampal neurons using confocal and TauSTED (stimulated emission depletion) microscopy. TauSTED is based on super-resolution STED nanoscopy, which combines high spatial resolution imaging (&amp;lt;40\u00a0nm) with fluorescence lifetime information, thus reducing background noise and improving image quality. We applied TauSTED imaging utilizing biotracker FerroFarRed Fe(II) and found that labile iron was present as submicrometric puncta in dendrites and axons. Some of these iron-rich structures are mobile and move along neuritic pathways, arguing for a labile iron transport mechanism in neurons. This super-resolution imaging approach offers a new perspective for studying the dynamic mechanisms of axonal and dendritic transport of iron at high spatial resolution in living neurons. In addition, this methodology could be transposed to the imaging of other fluorescent metal sensors.<\/jats:p>","DOI":"10.1093\/mtomcs\/mfad074","type":"journal-article","created":{"date-parts":[[2023,12,26]],"date-time":"2023-12-26T00:06:35Z","timestamp":1703549195000},"source":"Crossref","is-referenced-by-count":10,"title":["TauSTED super-resolution imaging of labile iron in primary hippocampal neurons"],"prefix":"10.1093","volume":"16","author":[{"given":"Aiyarin","family":"Kittilukkana","sequence":"first","affiliation":[{"name":"Chiang Mai University, Faculty of Associated Medical Sciences, Department of Radiologic Technology, Center of Excellence for Molecular Imaging (CEMI) , 50200 Chiang Mai , Thailand"},{"name":"University Bordeaux, CNRS, LP2I Bordeaux, UMR 5797 , Chemical Imaging and Speciation, F- 33170 Gradignan , France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9253-4581","authenticated-orcid":false,"given":"Asuncion","family":"Carmona","sequence":"additional","affiliation":[{"name":"University Bordeaux, CNRS, LP2I Bordeaux , UMR 5797, Chemical Imaging and Speciation, F- 33170 Gradignan , France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chalermchai","family":"Pilapong","sequence":"additional","affiliation":[{"name":"Chiang Mai University, Faculty of Associated Medical Sciences, Department of Radiologic Technology, Center of Excellence for Molecular Imaging (CEMI) , 50200 Chiang Mai , Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1692-5406","authenticated-orcid":false,"given":"Richard","family":"Ortega","sequence":"additional","affiliation":[{"name":"University Bordeaux, CNRS, LP2I Bordeaux , UMR 5797, Chemical Imaging and Speciation, F- 33170 Gradignan , France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,12,26]]},"reference":[{"key":"2024010515562770400_bib1","first-page":"164","article-title":"Review on Iron and Its Importance for Human Health","volume":"19","author":"Abbaspour","year":"2014","journal-title":"J. Res. Med. Sci."},{"issue":"4","key":"2024010515562770400_bib2","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1002\/(SICI)1097-4547(19981115)54:4<486::AID-JNR6>3.0.CO;2-I","article-title":"Evidence for Low Molecular Weight, Non-Transferrin-Bound Iron in Rat Brain and Cerebrospinal Fluid","volume":"54","author":"Moos","year":"1998","journal-title":"J. Neurosci. Res."},{"issue":"4","key":"2024010515562770400_bib3","doi-asserted-by":"crossref","first-page":"1109","DOI":"10.3233\/JAD-170097","article-title":"Transport of Non-Transferrin Bound Iron to the Brain: Implications for Alzheimer's Disease","volume":"58","author":"Tripathi","year":"2017","journal-title":"J. Alzheimers Dis."},{"issue":"8","key":"2024010515562770400_bib4","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.1016\/S0891-5849(02)01006-7","article-title":"The Labile Iron Pool: Characterization, Measurement, and Participation in Cellular Processes","volume":"33","author":"Kakhlon","year":"2002","journal-title":"Free Radical Biol. Med."},{"issue":"5","key":"2024010515562770400_bib5","doi-asserted-by":"crossref","first-page":"104691","DOI":"10.1016\/j.jbc.2023.104691","article-title":"PTEN-Induced Kinase PINK1 Supports Colorectal Cancer Growth by Regulating the Labile Iron Pool","volume":"299","author":"Chen","year":"2023","journal-title":"J. Biol. Chem."},{"issue":"1","key":"2024010515562770400_bib6","doi-asserted-by":"crossref","first-page":"51","DOI":"10.4155\/fmc.09.140","article-title":"Mechanisms of Brain Iron Transport: Insight into Neurodegeneration and CNS Disorders","volume":"2","author":"Mills","year":"2010","journal-title":"Future Med. Chem."},{"issue":"10","key":"2024010515562770400_bib7","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.1016\/S1474-4422(14)70117-6","article-title":"The Role of Iron in Brain Ageing and Neurodegenerative Disorders","volume":"13","author":"Ward","year":"2014","journal-title":"Lancet Neurol."},{"key":"2024010515562770400_bib8","doi-asserted-by":"crossref","first-page":"180","DOI":"10.3389\/fnins.2019.00180","article-title":"Iron in Neurodegeneration\u2014Cause or Consequence?","volume":"13","author":"Ndayisaba","year":"2019","journal-title":"Front. Neurosci."},{"issue":"5","key":"2024010515562770400_bib9","doi-asserted-by":"crossref","first-page":"668","DOI":"10.1111\/jnc.13040","article-title":"Molecular Mechanisms of Non-Transferrin-Bound and Transferring-Bound Iron Uptake in Primary Hippocampal Neurons","volume":"133","author":"Ji","year":"2015","journal-title":"J. Neurochem."},{"key":"2024010515562770400_bib10","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1016\/j.freeradbiomed.2015.03.033","article-title":"Role of Labile Iron in the Toxicity of Pharmacological Ascorbate","volume":"84","author":"Du","year":"2015","journal-title":"Free Radical Biol. Med."},{"issue":"2","key":"2024010515562770400_bib11","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1042\/BJ20061840","article-title":"Does the Calcein-AM Method Assay the Total Cellular \u2018Labile Iron Pool\u2019 or Only a Fraction of It?","volume":"403","author":"Tenopoulou","year":"2007","journal-title":"Biochem. J."},{"issue":"5","key":"2024010515562770400_bib12","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1267\/ahc.18015","article-title":"Development of Chemical Tools for Imaging of Fe(II) Ions in Living Cells: a Review","volume":"51","author":"Hirayama","year":"2018","journal-title":"Acta Histochem. Cytochem."},{"issue":"3","key":"2024010515562770400_bib13","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1016\/j.molcel.2020.01.003","article-title":"Maintaining Iron Homeostasis Is the Key Role of Lysosomal Acidity for Cell Proliferation","volume":"77","author":"Weber","year":"2020","journal-title":"Mol. Cell"},{"issue":"1","key":"2024010515562770400_bib14","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1111\/jnc.15583","article-title":"Heterogeneity of Ferrous Iron-Containing Endolysosomes and Effects of Endolysosome Iron on Endolysosome Numbers, Sizes, and Localization Patterns","volume":"161","author":"Halcrow","year":"2022","journal-title":"J. Neurochem."},{"issue":"1","key":"2024010515562770400_bib15","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1039\/C8MT00212F","article-title":"Organelle-Specific Analysis of Labile Fe(ii) during Ferroptosis by Using a Cocktail of Various Colour Organelle-Targeted Fluorescent Probes","volume":"11","author":"Hirayama","year":"2019","journal-title":"Metallomics"},{"issue":"3","key":"2024010515562770400_bib16","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1038\/nmeth.4593","article-title":"STED Super-Resolved Microscopy","volume":"15","author":"Vicidomini","year":"2018","journal-title":"Nat. Methods"},{"issue":"2","key":"2024010515562770400_bib17","doi-asserted-by":"crossref","first-page":"L14","DOI":"10.1529\/biophysj.107.120154","article-title":"The Phasor Approach to Fluorescence Lifetime Imaging Analysis","volume":"94","author":"Digman","year":"2008","journal-title":"Biophys. J."},{"issue":"5","key":"2024010515562770400_bib18","doi-asserted-by":"crossref","first-page":"2406","DOI":"10.1038\/nprot.2006.356","article-title":"Culturing Hippocampal Neurons","volume":"1","author":"Kaech","year":"2006","journal-title":"Nat. Protoc."},{"key":"2024010515562770400_bib19","article-title":"TauSTED: Pushing STED Beyond Its Limits with Lifetime","volume-title":"Leica Microsystems Application Note","author":"Alvarez","year":"2021"},{"issue":"10","key":"2024010515562770400_bib20","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.1038\/s41592-022-01588-y","article-title":"Event-Triggered STED Imaging","volume":"19","author":"Alvelid","year":"2022","journal-title":"Nat. Methods"},{"issue":"19","key":"2024010515562770400_bib21","doi-asserted-by":"crossref","first-page":"11971","DOI":"10.1021\/acs.chemrev.0c01174","article-title":"Super-Resolving Microscopy in Neuroscience","volume":"121","author":"Werner","year":"2021","journal-title":"Chem. Rev."},{"issue":"4","key":"2024010515562770400_bib22","doi-asserted-by":"crossref","first-page":"043701","DOI":"10.1063\/1.4799665","article-title":"Tuning Donut Profile for Spatial Resolution in Stimulated Emission Depletion Microscopy","volume":"84","author":"Neupane","year":"2013","journal-title":"Rev. Sci. Instrum."},{"issue":"8","key":"2024010515562770400_bib23","doi-asserted-by":"crossref","first-page":"886","DOI":"10.1016\/j.semcdb.2009.08.006","article-title":"Fluorescence Microscopy Below the Diffraction Limit","volume":"20","author":"Patterson","year":"2009","journal-title":"Semin. Cell Dev. Biol."},{"issue":"1","key":"2024010515562770400_bib24","doi-asserted-by":"crossref","first-page":"20782","DOI":"10.1038\/s41598-021-00301-x","article-title":"Evaluation of STED Super-Resolution Image Quality by Image Correlation Spectroscopy (QuICS)","volume":"11","author":"Cerutti","year":"2021","journal-title":"Sci. Rep."},{"issue":"4","key":"2024010515562770400_bib25","doi-asserted-by":"crossref","first-page":"2807","DOI":"10.1529\/biophysj.104.045492","article-title":"A Fast Global Fitting Algorithm for Fluorescence Lifetime Imaging Microscopy Based on Image Segmentation","volume":"87","author":"Pelet","year":"2004","journal-title":"Biophys. J."},{"issue":"1","key":"2024010515562770400_bib26","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1146\/annurev-biophys-062920-063631","article-title":"The Phasor Plot: a Universal Circle to Advance Fluorescence Lifetime Analysis and Interpretation","volume":"50","author":"Malacrida","year":"2021","journal-title":"Annu. Rev. Biophys."},{"issue":"7","key":"2024010515562770400_bib27","doi-asserted-by":"crossref","first-page":"7362","DOI":"10.1364\/OE.20.007362","article-title":"Strategies to Maximize the Performance of a STED Microscope","volume":"20","author":"Galiani","year":"2012","journal-title":"Opt. Express"},{"issue":"10","key":"2024010515562770400_bib28","doi-asserted-by":"crossref","first-page":"mfac064","DOI":"10.1093\/mtomcs\/mfac064","article-title":"Characterising Murine Hippocampal Iron Homeostasis, in Relation to Markers of Brain Inflammation and Metabolism, During Ageing","volume":"14","author":"Ellison","year":"2022","journal-title":"Metallomics"},{"issue":"12","key":"2024010515562770400_bib29","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1038\/s41589-019-0371-x","article-title":"Axonal Iron Transport in the Brain Modulates Anxiety-Related Behaviors","volume":"15","author":"Wang","year":"2019","journal-title":"Nat. Chem. Biol."},{"key":"2024010515562770400_bib30","doi-asserted-by":"crossref","first-page":"e62234","DOI":"10.7554\/eLife.62334","article-title":"Correlating STED and Synchrotron XRF Nano-Imaging Unveils Cosegregation of Metals and Cytoskeleton Proteins in Dendrites","volume":"9","author":"Domart","year":"2020","journal-title":"eLife"},{"key":"2024010515562770400_bib31","doi-asserted-by":"crossref","first-page":"109702","DOI":"10.1016\/j.jneumeth.2022.109702","article-title":"Correlative Nano-Imaging of Metals and Proteins in Primary Neurons by Synchrotron X-Ray Fluorescence and STED Super Resolution Microscopy: Experimental Validation","volume":"381","author":"Ortega","year":"2022","journal-title":"J. Neurosci. Methods"}],"container-title":["Metallomics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/metallomics\/advance-article-pdf\/doi\/10.1093\/mtomcs\/mfad074\/54879489\/mfad074.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/metallomics\/article-pdf\/16\/1\/mfad074\/55105756\/mfad074.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/metallomics\/article-pdf\/16\/1\/mfad074\/55105756\/mfad074.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,5]],"date-time":"2024-01-05T15:57:17Z","timestamp":1704470237000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/metallomics\/article\/doi\/10.1093\/mtomcs\/mfad074\/7499731"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,26]]},"references-count":31,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2024,1,5]]}},"URL":"https:\/\/doi.org\/10.1093\/mtomcs\/mfad074","relation":{},"ISSN":["1756-591X"],"issn-type":[{"value":"1756-591X","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2024,1]]},"published":{"date-parts":[[2023,12,26]]},"article-number":"mfad074"}}