{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T11:10:32Z","timestamp":1788779432407,"version":"build-2803163510"},"reference-count":46,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2014\/06870\u20100"],"award-info":[{"award-number":["2014\/06870\u20100"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2025\/00247\u20103"],"award-info":[{"award-number":["2025\/00247\u20103"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["304177\/2024\u20105"],"award-info":[{"award-number":["304177\/2024\u20105"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["001"],"award-info":[{"award-number":["001"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microscopy Res &amp;amp; Technique"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Mangrove ecosystems are frequently exposed to high concentrations of iron (Fe) in sediments, resulting in Fe accumulation in plant tissues. Although Fe is an essential micronutrient involved in several metabolic processes, its excess requires efficient mechanisms of compartmentalization and storage to maintain cellular homeostasis. Histochemical detection using the Perls reaction has usually been applied to identify ferric iron (Fe\n                    <jats:sup>3+<\/jats:sup>\n                    ) in biological tissues; however, the combination of this technique with ultrastructural and elemental analyses remains relatively unexplored in plant cells. In this study, we investigated Fe localization in leaf tissues of\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Rhizophora mangle<\/jats:italic>\n                    <\/jats:styled-content>\n                    L. (Rhizophoraceae), a dominant mangrove species, by combining complementary approaches, including Perls cytochemical reaction, transmission electron microscopy (TEM), scanning transmission electron microscopy coupled with high\u2010angle annular dark\u2010field imaging (STEM\u2010HAADF), and energy\u2010dispersive X\u2010ray spectroscopy (EDS). Perls\u2010positive electron\u2010dense deposits were visualized at the ultrastructural level, and their elemental composition was further characterized by EDS analyses. Fe\u2010containing deposits were detected in the epidermis, mesophyll parenchyma, mucilage cells, and vascular tissues, as well as in multiple cellular compartments, including plastids, mitochondria, vacuoles, cell walls, intercellular spaces, and plasmodesmata, whereas sclerenchyma cells showed no detectable Fe\u2010containing deposits. The combination of Perls reaction with TEM, STEM\u2010HAADF, and EDS provides a complementary approach for high\u2010resolution visualization and elemental characterization of Fe\u2010containing deposits at the subcellular level. This integrated methodology may facilitate the investigation of Fe distribution and compartmentalization in plant tissues under contrasting conditions of Fe availability.\n                  <\/jats:p>","DOI":"10.1002\/jemt.70180","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T10:28:51Z","timestamp":1788776931000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Subcellular Localization of Iron in\n                    <i>Rhizophora mangle<\/i>\n                    Leaves Revealed by Integrated Perls Reaction,\n                    <scp>TEM<\/scp>\n                    ,\n                    <scp>STEM<\/scp>\n                    \u2010\n                    <scp>HAADF<\/scp>\n                    , and\n                    <scp>EDS<\/scp>\n                    Analyses"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3137-8551","authenticated-orcid":false,"given":"Silvia Rodrigues","family":"Machado","sequence":"first","affiliation":[{"name":"Center of Electron Microscopy (CME) Institute of Biosciences of Botucatu (IBB), S\u00e3o Paulo State University (UNESP)  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7705-5251","authenticated-orcid":false,"given":"Hiulana Pereira","family":"Arrivabene","sequence":"additional","affiliation":[{"name":"Postgraduate Program in Plant Biology Institute of Biosciences of Botucatu (IBB), S\u00e3o Paulo State University (UNESP)  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9516-2031","authenticated-orcid":false,"given":"Camilla Rozindo Dias","family":"Milanez","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences Federal University of Esp\u00edrito Santo (UFES)  Vit\u00f3ria Esp\u00edrito Santo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0452-3701","authenticated-orcid":false,"given":"Margarida Juri","family":"Saeki","sequence":"additional","affiliation":[{"name":"Department of Chemistry &amp; Biochemistry Institute of Biosciences of Botucatu (IBB), S\u00e3o Paulo State University (UNESP)  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7516-9845","authenticated-orcid":false,"given":"Yve","family":"Canaveze","sequence":"additional","affiliation":[{"name":"Department of Botany Federal University of S\u00e3o Carlos (UFSCar)  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,7]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"crossref","first-page":"335e340","DOI":"10.1016\/j.plaphy.2007.03.020","article-title":"Composition, Speciation and Distribution of Iron Minerals in \n                     Imperata cylindrica","volume":"45","author":"Amils R.","year":"2007","journal-title":"Plant Physiology and Biochemistry"},{"key":"e_1_2_11_3_1","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1007\/BF01033631","article-title":"Imidazole\u2010Buffered Osmium Tetroxide: An Excellent Stain for Visualization of Lipids in Transmission Electron Microscopy","volume":"75","author":"Angerm\u00fcller S.","year":"1982","journal-title":"Histochemistry"},{"key":"e_1_2_11_4_1","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1557\/mrs.2011.329","article-title":"Development of Electron Energy\u2010Loss Spectroscopy in the Biological Sciences","volume":"37","author":"Aronova M. A.","year":"2012","journal-title":"MRS Bulletin"},{"key":"e_1_2_11_5_1","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1016\/j.envpol.2016.05.019","article-title":"Differential Bioaccumulation and Translocation Patterns in Three Mangrove Plants Experimentally Exposed to Iron. Consequences for Environmental Sensing","volume":"215","author":"Arrivabene H. P.","year":"2016","journal-title":"Environmental Pollution"},{"key":"e_1_2_11_6_1","doi-asserted-by":"crossref","first-page":"27e34","DOI":"10.1016\/j.chemosphere.2015.01.011","article-title":"Effect of Pollution by Particulate Iron on the Morphoanatomy, Histochemistry, and Bioaccumulation of Three Mangrove Plant Species in Brazil","volume":"127","author":"Arrivabene H. P.","year":"2015","journal-title":"Chemosphere"},{"key":"e_1_2_11_7_1","volume-title":"Theory and Practice of Histological Techniques","author":"Bancroft J. D.","year":"1996"},{"key":"e_1_2_11_8_1","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1039\/C7MT00136C","article-title":"Iron Homeostasis in Plants \u2010 a Brief Overview","volume":"9","author":"Connorton J. M.","year":"2017","journal-title":"Metallomics"},{"key":"e_1_2_11_9_1","doi-asserted-by":"crossref","first-page":"152794","DOI":"10.1016\/j.flora.2025.152794","article-title":"Iron Histolocalization in Amazon Canga Plants Reveals Phylogeny\u2010Driven Adaptation Strategies","volume":"330","author":"Filgueira J. P. P. S.","year":"2025","journal-title":"Flora"},{"key":"e_1_2_11_10_1","doi-asserted-by":"crossref","first-page":"3351","DOI":"10.1038\/s41467-023-38880-0","article-title":"Cell Type\u2010Specific Mapping of Ion Distribution in \n                     Arabidopsis thaliana\n                   Roots","volume":"14","author":"Giehl R. F. H.","year":"2023","journal-title":"Nature Communications"},{"key":"e_1_2_11_11_1","doi-asserted-by":"crossref","DOI":"10.1007\/978-1-4939-6676-9","volume-title":"Scanning Electron Microscopy and X\u2010Ray Microanalysis","author":"Goldstein J.","year":"2018"},{"key":"e_1_2_11_12_1","doi-asserted-by":"crossref","first-page":"111124","DOI":"10.1016\/j.plaphy.2026.111124","article-title":"Ferritin\u2010Mediated Transient Iron Sequestration Facilitates Chloroplast Iron Recycling During Leaf Senescence","volume":"232","author":"Gracheva M.","year":"2026","journal-title":"Plant Physiology and Biochemistry"},{"key":"e_1_2_11_13_1","doi-asserted-by":"crossref","first-page":"2523","DOI":"10.1104\/pp.104.045633","article-title":"FRD3 Controls Iron Localization in Arabidopsis","volume":"136","author":"Green L. S.","year":"2004","journal-title":"Plant Physiology"},{"key":"e_1_2_11_14_1","volume-title":"Principles and Techniques of Electron Microscopy: Biological Applications","author":"Hayat M. A.","year":"2000"},{"key":"e_1_2_11_15_1","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/j.jenvman.2016.04.011","article-title":"Application of Mucilage From Dicerocaryum eriocarpum Plant as Biosorption Medium in the Removal of Selected Heavy Metal Ions","volume":"177","author":"Jones B. O.","year":"2016","journal-title":"Journal of Environmental Management"},{"key":"e_1_2_11_16_1","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1126\/science.1132563","article-title":"Localization of Iron in Arabidopsis Seed Requires the Vacuolar Membrane Transporter VIT1","volume":"314","author":"Kim S. A.","year":"2006","journal-title":"Science"},{"key":"e_1_2_11_17_1","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1093\/jmicro\/dfac018","article-title":"Improved Chemical Fixation of Lipid\u2010Secreting Plant Cells for Transmission Electron Microscopy","volume":"71","author":"Kiyoto S.","year":"2022","journal-title":"Microscopy"},{"key":"e_1_2_11_18_1","doi-asserted-by":"crossref","first-page":"900","DOI":"10.2183\/pjab.86.900","article-title":"Recent Insights Into Iron Homeostasis and Their Application in Graminaceous Crops","volume":"86","author":"Kobayashi T.","year":"2010","journal-title":"Proceedings of the Japan Academy. Series B, Physical and Biological Sciences"},{"key":"e_1_2_11_19_1","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1083\/jcb.34.2.627","article-title":"A Chromatographic and Spectrophotometric Study of the Products of the Reaction of Osmium Tetroxide With Unsaturated Lipids","volume":"34","author":"Korn E. D.","year":"1967","journal-title":"Journal of Cell Biology"},{"key":"e_1_2_11_20_1","doi-asserted-by":"crossref","first-page":"3395","DOI":"10.3390\/ijms21093395","article-title":"Regulation of Iron Homeostasis and Use in Chloroplasts","volume":"21","author":"Kroh G. E.","year":"2020","journal-title":"International Journal of Molecular Sciences"},{"key":"e_1_2_11_21_1","doi-asserted-by":"crossref","first-page":"4041","DOI":"10.1038\/sj.emboj.7600864","article-title":"Mobilization of Vacuolar Iron by AtNRAMP3 and AtNRAMP4 Is Essential for Seed Germination on Low Iron","volume":"24","author":"Lanquar V.","year":"2005","journal-title":"EMBO Journal"},{"key":"e_1_2_11_22_1","first-page":"300","article-title":"Novel Techniques in Electron Microscopy","volume":"14","author":"Leapman R. D.","year":"2003","journal-title":"Current Opinion in Biotechnology"},{"key":"e_1_2_11_23_1","doi-asserted-by":"crossref","first-page":"e14","DOI":"10.1017\/qpb.2025.11","article-title":"Untangling Iron Threads: A Deep Dive Into Plant Intracellular Pools","volume":"6","author":"Leskova A.","year":"2025","journal-title":"Quantitative Plant Biology"},{"key":"e_1_2_11_24_1","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1186\/s12284-025-00816-1","article-title":"Facilitating Phloem\u2010Mediated Iron Transport Can Improve the Adaptation of Rice Seedlings to Iron Deficiency Stress","volume":"18","author":"Lin Y.","year":"2025","journal-title":"Rice"},{"key":"e_1_2_11_25_1","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1016\/j.tplants.2023.03.007","article-title":"Beyond Iron\u2010Storage Pool: Functions of Plant Apoplastic Iron During Stress","volume":"28","author":"Liu X. X.","year":"2023","journal-title":"Trends in Plant Science"},{"key":"e_1_2_11_26_1","doi-asserted-by":"crossref","first-page":"2751","DOI":"10.4081\/ejh.2016.2751","article-title":"Transmission Electron Microscopy for Nanomedicine: Novel Applications for Long\u2010Established Techniques","volume":"60","author":"Malatesta M.","year":"2016","journal-title":"European Journal of Histochemistry"},{"key":"e_1_2_11_27_1","doi-asserted-by":"crossref","first-page":"12789","DOI":"10.3390\/ijms222312789","article-title":"Transmission Electron Microscopy as a Powerful Tool to Investigate the Interaction of Nanoparticles With Subcellular Structures","volume":"22","author":"Malatesta M.","year":"2021","journal-title":"International Journal of Molecular Sciences"},{"key":"e_1_2_11_28_1","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.foodhyd.2017.03.020","article-title":"Chemical Structure and Physical\u2010Chemical Properties of Mucilage From the Leaves of \n                     Pereskia aculeata","volume":"70","author":"Martin A. A.","year":"2017","journal-title":"Food Hydrocolloids"},{"key":"e_1_2_11_29_1","first-page":"233","article-title":"Cellular and Subcellular Localization of Non\u2010Heme Iron Using the Perls' Prussian Blue Reaction","volume":"127","author":"Meguro R.","year":"2007","journal-title":"Histochemistry and Cell Biology"},{"key":"e_1_2_11_30_1","doi-asserted-by":"crossref","first-page":"100344","DOI":"10.1016\/j.rhisph.2021.100344","article-title":"Plant Mucilage Components and Their Functions in the Rhizosphere","volume":"18","author":"Nazari M.","year":"2021","journal-title":"Rhizosphere"},{"key":"e_1_2_11_31_1","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1016\/0272-7714(90)90017-L","article-title":"Factors Affecting the Hydrochemistry of a Mangrove Tidal Creek, Sepetiba Bay, Brazil","volume":"31","author":"Ovalle A. R. C.","year":"1990","journal-title":"Estuarine, Coastal and Shelf Science"},{"key":"e_1_2_11_32_1","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1134\/S1021443707020136","article-title":"Ultrastructure of Ferritin in the Leaves of \n                     Mesembryanthemum crystallinum\n                   Under Stress Conditions","volume":"54","author":"Paramonova N. V.","year":"2007","journal-title":"Russian Journal of Plant Physiology"},{"key":"e_1_2_11_33_1","first-page":"707","article-title":"Ultrastructural Cytochemistry of Iron Absorption","volume":"93","author":"Parmley R. T.","year":"1978","journal-title":"American Journal of Pathology"},{"key":"e_1_2_11_34_1","doi-asserted-by":"crossref","DOI":"10.1007\/978-1-4419-7200-2","volume-title":"Scanning Transmission Electron Microscopy: Imaging and Analysis","author":"Pennycook S. J.","year":"2011"},{"key":"e_1_2_11_35_1","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1007\/BF01282540","article-title":"Iron\u2010Rich Particles in Embryos of Seeds From the Family Pinaceae","volume":"202","author":"Reid D. A.","year":"1998","journal-title":"Protoplasma"},{"key":"e_1_2_11_36_1","doi-asserted-by":"crossref","first-page":"781e789","DOI":"10.1111\/j.1469-8137.2004.01264.x","article-title":"Internal Iron Biomineralization in \n                     Imperata cylindrica\n                  , a Perennial Grass: Chemical Composition, Speciation and Plant Localization","volume":"165","author":"Rodr\u00edguez N.","year":"2005","journal-title":"New Phytologist"},{"key":"e_1_2_11_37_1","first-page":"350","article-title":"New Insights Into Fe Localization in Plant Tissues","volume":"4","author":"Roschzttardtz H.","year":"2013","journal-title":"Frontiers in Plant Science, Plant Nutrition"},{"key":"e_1_2_11_38_1","doi-asserted-by":"crossref","first-page":"1717","DOI":"10.1093\/jxb\/erac030","article-title":"Iron in Leaves: Chemical Forms, Signaling, and In\u2010Cell Distribution","volume":"73","author":"S\u00e1gi\u2010Kaz\u00e1r M.","year":"2022","journal-title":"Journal of Experimental Botany"},{"key":"e_1_2_11_39_1","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/S0022-5320(72)80007-8","article-title":"Electron Microscopical Observations of Leaf Ferritin From Iron\u2010Treated Xanthium Plants: Localization and Diversity in the Organelle","volume":"39","author":"Seckbach J. J.","year":"1972","journal-title":"Journal of Ultrastructure and Molecular Structure Research"},{"key":"e_1_2_11_40_1","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1080\/01904168209362966","article-title":"Ferreting Out the Secrets of Plant Ferritin: A Review","volume":"5","author":"Seckbach J. J.","year":"1982","journal-title":"Journal of Plant Nutrition"},{"key":"e_1_2_11_41_1","doi-asserted-by":"crossref","first-page":"1644","DOI":"10.1111\/nph.17440","article-title":"Subcellular Dynamics Studies of Iron Reveal How Tissue\u2010Specific Distribution Patterns Are Established in Developing Wheat Grains","volume":"231","author":"Sheraz S.","year":"2021","journal-title":"New Phytologist"},{"key":"e_1_2_11_42_1","first-page":"100627","article-title":"Perls' Prussian Blue Staining and Chemistry of Prussian Blue and Turnbull Blue","volume":"11","author":"Sonoda A.","year":"2025","journal-title":"Forensic Science International: Synergy"},{"key":"e_1_2_11_43_1","volume-title":"Plant Physiology and Development","author":"Taiz L.","year":"2022"},{"key":"e_1_2_11_44_1","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1146\/annurev.bi.56.070187.001445","article-title":"Ferritin: Structure, Gene Regulation, and Cellular Function in Animals, Plants and Microorganisms","volume":"56","author":"Theil E. C.","year":"1987","journal-title":"Annual Review of Biochemistry"},{"key":"e_1_2_11_45_1","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.3390\/polym13071066","article-title":"Comprehensive Review on Plant\u2010Derived Mucilage: Characterization, Functional Properties, Applications, and Its Utilization for Nanocarrier Fabrication","volume":"13","author":"Tosif M. M.","year":"2021","journal-title":"Polymers"},{"key":"e_1_2_11_46_1","doi-asserted-by":"crossref","DOI":"10.1007\/978-0-387-76501-3","volume-title":"Transmission Electron Microscopy: A Textbook for Materials Science","author":"Williams D. B.","year":"2009"},{"key":"e_1_2_11_47_1","doi-asserted-by":"crossref","first-page":"5557","DOI":"10.1093\/jxb\/erae307","article-title":"New Insights Into Plasmodesmata: Complex \u2018Protoplasmic Connecting Threads\u2019","volume":"75","author":"Zanini A. A.","year":"2024","journal-title":"Journal of Experimental Botany"}],"container-title":["Microscopy Research and Technique"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jemt.70180","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jemt.70180","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jemt.70180","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T10:28:58Z","timestamp":1788776938000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/jemt.70180"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,7]]},"references-count":46,"alternative-id":["10.1002\/jemt.70180"],"URL":"https:\/\/doi.org\/10.1002\/jemt.70180","archive":["Portico"],"relation":{},"ISSN":["1059-910X","1097-0029"],"issn-type":[{"value":"1059-910X","type":"print"},{"value":"1097-0029","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,7]]},"assertion":[{"value":"2026-04-07","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-31","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-07","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"jemt.70180"}}