Three-dimensional structure of entire hydrated murine hearts at histological resolution
Abstract Imaging the entire cardiomyocyte network in entire small animal hearts at single cell resolution is a formidable challenge. Optical microscopy provides sufficient contrast and resolution in 2d, however fails to deliver non-destructive 3d reconstructions with isotropic resolution. It require...
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Nature Portfolio
2025-01-01
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| Online Access: | https://doi.org/10.1038/s41598-024-83853-y |
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| author | Jasper Frohn Frederik Böddeker Marius Reichardt Hendrik Bruns Titus Czajka Amara Khan Ludovic Broche Michael Krisch Alberto Bravin Frauke Alves Jana Zschüntzsch Tim Salditt |
| author_facet | Jasper Frohn Frederik Böddeker Marius Reichardt Hendrik Bruns Titus Czajka Amara Khan Ludovic Broche Michael Krisch Alberto Bravin Frauke Alves Jana Zschüntzsch Tim Salditt |
| author_sort | Jasper Frohn |
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| description | Abstract Imaging the entire cardiomyocyte network in entire small animal hearts at single cell resolution is a formidable challenge. Optical microscopy provides sufficient contrast and resolution in 2d, however fails to deliver non-destructive 3d reconstructions with isotropic resolution. It requires several invasive preparation steps, which introduce structural artefacts, namely dehydration, physical slicing and staining, or for the case of light sheet microscopy also clearing of the tissue. Our goal is to provide 3d reconstructions of the cardiomyocyte network in entire hydrated murine hearts, and to develop a methodology for quantitative analysis of heart pathologies based on X-ray phase contrast computed tomography (XPCT). We have used XPCT at two beamlines of the extremely brilliant source (EBS) at the European Synchrotron Radiation Facility (ESRF) to scan wild-type murine hearts at high resolution, as well as a series of murine hearts of different pathological models, at reduced resolution and higher throughput. All hearts were obtained from the small animal facility of the university medical center in Göttingen. The hearts were fixed in formalin, stored and measured non-destructively in phosphate buffer solution. The high resolution dataset allows to discern individual cardiomyocytes in the tissue. All datasets have been analyzed using semi-automated image segmentation of the ventricles, rotation into a common coordinate system, classification into different anatomical compartments, and finally the structure tensor approach. A 3d streamline representation of the cardiomyocyte orientation vector field is provided. The different cardiovascular disease models are analysed based on metrics derived from the 3d structure tensor. An entire hydrated murine heart has been covered at an isotropic voxel size of 1.6 $$\upmu$$ μ m (distributed over several volumes). A binned and fused dataset of this heart is available at 3.2 $$\upmu$$ μ m, and has been analyzed by the structure tensor approach to yield the ventricular cardiomyocyte network or mesh, i.e. the aggregation of the cardiomyocyte chains in particular in the ventricular wall. Semi-automatic determination of structural metrics is already achieved and the corresponding tools and resulting data are made publically available. XPCT using extremely brilliant undulator radiation is close to achieve single cell reconstruction in an entire small animal organ. |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-01-01 |
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| spelling | doaj-art-7aaf3f2fba824dbbb85cd6ee676927c92025-01-26T12:25:35ZengNature PortfolioScientific Reports2045-23222025-01-0115111610.1038/s41598-024-83853-yThree-dimensional structure of entire hydrated murine hearts at histological resolutionJasper Frohn0Frederik Böddeker1Marius Reichardt2Hendrik Bruns3Titus Czajka4Amara Khan5Ludovic Broche6Michael Krisch7Alberto Bravin8Frauke Alves9Jana Zschüntzsch10Tim Salditt11Institute for X-ray Physics, Georg-August University GöttingenInstitute for X-ray Physics, Georg-August University GöttingenInstitute for X-ray Physics, Georg-August University GöttingenInstitute for X-ray Physics, Georg-August University GöttingenInstitute for X-ray Physics, Georg-August University GöttingenGeorg-August University Göttingen, University Medical Center GöttingenMultiscale Bioimaging: From Molecular Machines to Networks of Excitable Cells (MBExC)Multiscale Bioimaging: From Molecular Machines to Networks of Excitable Cells (MBExC)Multiscale Bioimaging: From Molecular Machines to Networks of Excitable Cells (MBExC)Georg-August University Göttingen, University Medical Center GöttingenGeorg-August University Göttingen, University Medical Center GöttingenInstitute for X-ray Physics, Georg-August University GöttingenAbstract Imaging the entire cardiomyocyte network in entire small animal hearts at single cell resolution is a formidable challenge. Optical microscopy provides sufficient contrast and resolution in 2d, however fails to deliver non-destructive 3d reconstructions with isotropic resolution. It requires several invasive preparation steps, which introduce structural artefacts, namely dehydration, physical slicing and staining, or for the case of light sheet microscopy also clearing of the tissue. Our goal is to provide 3d reconstructions of the cardiomyocyte network in entire hydrated murine hearts, and to develop a methodology for quantitative analysis of heart pathologies based on X-ray phase contrast computed tomography (XPCT). We have used XPCT at two beamlines of the extremely brilliant source (EBS) at the European Synchrotron Radiation Facility (ESRF) to scan wild-type murine hearts at high resolution, as well as a series of murine hearts of different pathological models, at reduced resolution and higher throughput. All hearts were obtained from the small animal facility of the university medical center in Göttingen. The hearts were fixed in formalin, stored and measured non-destructively in phosphate buffer solution. The high resolution dataset allows to discern individual cardiomyocytes in the tissue. All datasets have been analyzed using semi-automated image segmentation of the ventricles, rotation into a common coordinate system, classification into different anatomical compartments, and finally the structure tensor approach. A 3d streamline representation of the cardiomyocyte orientation vector field is provided. The different cardiovascular disease models are analysed based on metrics derived from the 3d structure tensor. An entire hydrated murine heart has been covered at an isotropic voxel size of 1.6 $$\upmu$$ μ m (distributed over several volumes). A binned and fused dataset of this heart is available at 3.2 $$\upmu$$ μ m, and has been analyzed by the structure tensor approach to yield the ventricular cardiomyocyte network or mesh, i.e. the aggregation of the cardiomyocyte chains in particular in the ventricular wall. Semi-automatic determination of structural metrics is already achieved and the corresponding tools and resulting data are made publically available. XPCT using extremely brilliant undulator radiation is close to achieve single cell reconstruction in an entire small animal organ.https://doi.org/10.1038/s41598-024-83853-yVirtual 3d histologyCardiomyocyte networkX-ray phase-contrastX-ray tomographyOrgan imaging |
| spellingShingle | Jasper Frohn Frederik Böddeker Marius Reichardt Hendrik Bruns Titus Czajka Amara Khan Ludovic Broche Michael Krisch Alberto Bravin Frauke Alves Jana Zschüntzsch Tim Salditt Three-dimensional structure of entire hydrated murine hearts at histological resolution Scientific Reports Virtual 3d histology Cardiomyocyte network X-ray phase-contrast X-ray tomography Organ imaging |
| title | Three-dimensional structure of entire hydrated murine hearts at histological resolution |
| title_full | Three-dimensional structure of entire hydrated murine hearts at histological resolution |
| title_fullStr | Three-dimensional structure of entire hydrated murine hearts at histological resolution |
| title_full_unstemmed | Three-dimensional structure of entire hydrated murine hearts at histological resolution |
| title_short | Three-dimensional structure of entire hydrated murine hearts at histological resolution |
| title_sort | three dimensional structure of entire hydrated murine hearts at histological resolution |
| topic | Virtual 3d histology Cardiomyocyte network X-ray phase-contrast X-ray tomography Organ imaging |
| url | https://doi.org/10.1038/s41598-024-83853-y |
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