{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T20:09:58Z","timestamp":1789157398130,"version":"build-2803163510"},"reference-count":61,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T00:00:00Z","timestamp":1789084800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T00:00:00Z","timestamp":1789084800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["502822458"],"award-info":[{"award-number":["502822458"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["physoc.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Physiology"],"abstract":"<jats:sec>\n                    <jats:title>Abstract<\/jats:title>\n                    <jats:p>\n                      Understanding the three\u2010dimensional organisation of the myocardium requires imaging approaches that can resolve cellular structures across the entire heart. Existing methods are limited by trade\u2010offs between spatial resolution, imaging depth and throughput. Here we present a high\u2010throughput imaging pipeline for high\u2010fidelity reconstruction of whole mouse hearts. The approach combines a CUBIC tissue clearing protocol optimized for cardiac tissue, a dual\u2010camera selective plane\u2010illumination microscope (dual\u2010mesoSPIM) and an automated fusion algorithm that selects the highest\u2010contrast information from the opposing detection views. Whole hearts (\n                      <jats:italic>n<\/jats:italic>\n                      \u00a0=\u00a09) were imaged label free (using myocardial autofluorescence) at a voxel size of 3.25 \u00d7 3.25 \u00d7 3 \u00b5m\n                      <jats:sup>3<\/jats:sup>\n                      in approximately 25\u00a0min per sample. Dual\u2010view fusion significantly reduces depth\u2010dependent information loss compared to single\u2010camera acquisitions, enabling robust three\u2010dimensional reconstruction and morphometry of cardiomyocyte orientations and laminar tissue organisation across the ventricular walls. Quantitative morphometry of fibre and sheet orientations confirmed the expected transmural organisation of the myocardium, including the progressive rotation of the helix angle and the regional variation in sheet architecture. The protocol is compatible with endogenous fluorescent reporters and whole\u2010mount staining, allowing, for example, the reconstruction of the cardiac sympathetic nervous system throughout the intact mouse organ. Our new pipeline provides a rapid and scalable strategy for mesoscale mapping of cardiac cytoarchitecture in physiological and pathological models.\n                      <jats:boxed-text content-type=\"graphic\" position=\"anchor\">\n                        <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/tjp70782-gra-0001-m.png\">\n                          <jats:alt-text>image<\/jats:alt-text>\n                        <\/jats:graphic>\n                      <\/jats:boxed-text>\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Key points<\/jats:title>\n                    <jats:p>\n                      <jats:list list-type=\"bullet\">\n                        <jats:list-item>\n                          <jats:p>Understanding cardiac function requires mapping the three\u2010dimensional arrangement of cardiomyocytes throughout the whole organ, yet reconstructing this cytoarchitecture at cellular resolution across an intact heart remains technically demanding.<\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>We combined CUBIC tissue clearing with a dual\u2010camera light\u2010sheet microscope (dual\u2010mesoSPIM) to reconstruct the entire adult mouse heart in three dimensions at cellular scale, rapidly and with high fidelity.<\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>Acquiring and fusing images from two opposing cameras substantially reduced the loss of information at depth, allowing the local orientation of cardiomyocytes and myocardial sheetlets to be quantified throughout the ventricular walls, including the expected transmural rotation of the fibre helix angle.<\/jats:p>\n                        <\/jats:list-item>\n                        <jats:list-item>\n                          <jats:p>The approach preserves endogenous fluorescent reporters and is compatible with whole\u2010heart immunostaining, which we used to reconstruct the cardiac sympathetic innervation across the entire organ, opening the way to mapping cellular organization and its remodelling in health and disease.<\/jats:p>\n                        <\/jats:list-item>\n                      <\/jats:list>\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1113\/jp290463","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T19:30:18Z","timestamp":1789155018000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["High\u2010throughput, high\u2010fidelity reconstruction of whole mouse\u2010heart cytoarchitecture at cellular scale"],"prefix":"10.1113","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8301-2864","authenticated-orcid":false,"given":"Francesco","family":"Giardini","sequence":"first","affiliation":[{"name":"Institute of Clinical Physiology National Research Council (IFC\u2010CNR)  Florence Italy"},{"name":"Institute for Experimental Cardiovascular Medicine University Heart Center Freiburg \u2012 Bad KrozingenMedical Faculty and Medical Center \u2013 University of Freiburg  Freiburg im Breisgau Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Palandri","sequence":"additional","affiliation":[{"name":"National Institute of Optics National Research Council  Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sofia","family":"Botti","sequence":"additional","affiliation":[{"name":"MOX, Department of Mathematics Politecnico di Milano  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-8391-571X","authenticated-orcid":false,"given":"Camilla","family":"Olianti","sequence":"additional","affiliation":[{"name":"School of Cardiovascular and Metabolic Medicine &amp; Sciences and British Heart Foundation Centre of Research Excellence King's College London  London UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paolo","family":"Mele","sequence":"additional","affiliation":[{"name":"Department of Information Engineering University of Florence  Florence Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-7593-5014","authenticated-orcid":false,"given":"Niccol\u00f2","family":"Ballerini","sequence":"additional","affiliation":[{"name":"Department of Information Engineering University of Florence  Florence Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1239-8031","authenticated-orcid":false,"given":"Roberto","family":"Piersanti","sequence":"additional","affiliation":[{"name":"Department of Theoretical and Applied Science (DiSTA) eCampus University  Novedrate Italy"},{"name":"SMARTEST Research Center eCampus University  Novedrate Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mauro","family":"Giacca","sequence":"additional","affiliation":[{"name":"School of Cardiovascular and Metabolic Medicine &amp; Sciences and British Heart Foundation Centre of Research Excellence King's College London  London UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0416-6270","authenticated-orcid":false,"given":"Peter","family":"Kohl","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine University Heart Center Freiburg \u2012 Bad KrozingenMedical Faculty and Medical Center \u2013 University of Freiburg  Freiburg im Breisgau Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6773-9884","authenticated-orcid":false,"given":"Callum M.","family":"Zgierski\u2010Johnston","sequence":"additional","affiliation":[{"name":"Institute for Experimental Cardiovascular Medicine University Heart Center Freiburg \u2012 Bad KrozingenMedical Faculty and Medical Center \u2013 University of Freiburg  Freiburg im Breisgau Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7839-1361","authenticated-orcid":false,"given":"Elisabetta","family":"Cerbai","sequence":"additional","affiliation":[{"name":"Department of Neurosciences, Psychology, Drug Research and Child Health University of Florence  Florence Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4207-1400","authenticated-orcid":false,"given":"Francesco","family":"Regazzoni","sequence":"additional","affiliation":[{"name":"MOX, Department of Mathematics Politecnico di Milano  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9320-5085","authenticated-orcid":false,"given":"Leonardo","family":"Sacconi","sequence":"additional","affiliation":[{"name":"Institute of Clinical Physiology National Research Council (IFC\u2010CNR)  Florence Italy"},{"name":"Institute for Experimental Cardiovascular Medicine University Heart Center Freiburg \u2012 Bad KrozingenMedical Faculty and Medical Center \u2013 University of Freiburg  Freiburg im Breisgau Germany"},{"name":"European Laboratory for Non\u2010Linear Spectroscopy \u2013 LENS  Sesto Fiorentino Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,11]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587\u2010025\u201002882\u20108"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12968-014-0077-x"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btx180"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.media.2021.102265"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcvm.2022.945198"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvaf209"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2022113288"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/cnm.3185"},{"key":"e_1_2_6_10_1","doi-asserted-by":"crossref","unstructured":"Fern\u00e1ndez M. C. W\u00fclfers E. M. Madl J. Feliz S. P. Kohl P. Zgierski\u2010Johnston C. M. &Schneider\u2010Warme F.(2023).3D structure of fibroblasts and macrophages in the healthy and cryo\u2010ablated heart.biorxiv.https:\/\/www.biorxiv.org\/content\/10.1101\/2023.11.30.569388v1","DOI":"10.1101\/2023.11.30.569388"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.119.311148"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-024-83853-y"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.3390\/biology13020105"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/s44161-025-00728-9"},{"key":"e_1_2_6_15_1","article-title":"Mesoscopic optical imaging of whole mouse heart","volume":"176","author":"Giardini F.","year":"2021","journal-title":"Journal of Visualized Experiments"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1093\/ehjci\/jew314"},{"key":"e_1_2_6_17_1","unstructured":"Guastamacchia C. Piersanti R. Giardini F. Coppini R. Ferrantini C. Dede\u2019 L. Sacconi L. &Regazzoni F.(2026).The functional impact of myofiber macroscopic organization and disarray in computational models of the murine heart. arXiv.https:\/\/doi.org\/10.48550\/arXiv.2604.00881"},{"issue":"4","key":"e_1_2_6_18_1","first-page":"747","article-title":"Desmoplakin cardiomyopathy: Gene dose\u2010dependent myocardial remodeling, arrhythmias, and premature death","volume":"12","author":"Guazzo A.","year":"2026","journal-title":"Journal of the American College of Cardiology Clinical Electrophysiology"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/mrm.20622"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41569-019-0221-2"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0501-0"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.1998.274.5.H1627"},{"issue":"4","key":"e_1_2_6_23_1","first-page":"117:1","article-title":"Fast tetrahedral meshing in the wild","volume":"39","author":"Hu Y.","year":"2020","journal-title":"Association for Computing Machinery Transactions on Graphics"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jelectrocard.2005.06.096"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1109\/TMI.2011.2172690"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.1995.269.2.H571"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2797-4"},{"key":"e_1_2_6_28_1","doi-asserted-by":"crossref","unstructured":"Liu Y. M\u00fcller G. F. Kowitz L. Chobola T. Weiss K. Maier P. Luo J. Roe\u00dfing M. Stenzel M. Gr\u00fcneboom A. Paetzold J. Ert\u00fcrk A. Navab N. Marr C. Chen J. Huisken J. &Peng T.(2025).Leonardo: A toolset to correct sample\u2010induced artifacts in light sheet microscopy images. 2025.10.26.684661. bioRxiv.https:\/\/www.biorxiv.org\/content\/10.1101\/2025.10.26.684661v1","DOI":"10.1101\/2025.10.26.684661"},{"key":"e_1_2_6_29_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-019-0240-9"},{"key":"e_1_2_6_30_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.123.322791"},{"key":"e_1_2_6_31_1","doi-asserted-by":"publisher","DOI":"10.1364\/BOE.583082"},{"key":"e_1_2_6_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.softx.2020.100454"},{"key":"e_1_2_6_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yjmcc.2013.10.018"},{"key":"e_1_2_6_34_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.114.300507"},{"key":"e_1_2_6_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pbiomolbio.2021.07.012"},{"key":"e_1_2_6_36_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.4281"},{"key":"e_1_2_6_37_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCEP.116.004133"},{"issue":"3","key":"e_1_2_6_38_1","first-page":"408","article-title":"Excessive trabeculation of the left ventricle","volume":"16","author":"Petersen S. E.","year":"2023","journal-title":"Journal of the American College of Cardiology Cardiovasc Imaging"},{"key":"e_1_2_6_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cma.2020.113468"},{"key":"e_1_2_6_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmpb.2021.106223"},{"key":"e_1_2_6_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4224"},{"key":"e_1_2_6_42_1","doi-asserted-by":"publisher","DOI":"10.1038\/s43586-021-00080-9"},{"key":"e_1_2_6_43_1","doi-asserted-by":"publisher","DOI":"10.1002\/jemt.20200"},{"key":"e_1_2_6_44_1","doi-asserted-by":"publisher","DOI":"10.1038\/s44161-025-00623-3"},{"key":"e_1_2_6_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacc.2006.08.030"},{"key":"e_1_2_6_46_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.125.326291"},{"key":"e_1_2_6_47_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.00184.2023"},{"key":"e_1_2_6_48_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2010.09.016"},{"key":"e_1_2_6_49_1","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.24.3.339"},{"key":"e_1_2_6_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.03.042"},{"key":"e_1_2_6_51_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-cellbio-111315-125001"},{"key":"e_1_2_6_52_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep30573"},{"key":"e_1_2_6_53_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2023.08.007"},{"key":"e_1_2_6_54_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2014.123"},{"key":"e_1_2_6_55_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-46770-2"},{"key":"e_1_2_6_56_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0554-0"},{"key":"e_1_2_6_57_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-021-01317-x"},{"key":"e_1_2_6_58_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-024-65371-z"},{"key":"e_1_2_6_59_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.adh9925"},{"key":"e_1_2_6_60_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.00059.2022"},{"key":"e_1_2_6_61_1","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvs320"},{"key":"e_1_2_6_62_1","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.125.325580"}],"container-title":["The Journal of Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/physoc.onlinelibrary.wiley.com\/doi\/pdf\/10.1113\/JP290463","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/physoc.onlinelibrary.wiley.com\/doi\/full-xml\/10.1113\/JP290463","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/physoc.onlinelibrary.wiley.com\/doi\/pdf\/10.1113\/JP290463","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T19:30:21Z","timestamp":1789155021000},"score":1,"resource":{"primary":{"URL":"https:\/\/physoc.onlinelibrary.wiley.com\/doi\/10.1113\/JP290463"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,11]]},"references-count":61,"alternative-id":["10.1113\/JP290463"],"URL":"https:\/\/doi.org\/10.1113\/jp290463","archive":["Portico"],"relation":{},"ISSN":["0022-3751","1469-7793"],"issn-type":[{"value":"0022-3751","type":"print"},{"value":"1469-7793","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,11]]},"assertion":[{"value":"2026-04-14","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-08-04","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-11","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"JP290463"}}