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Phase-sensitive computed tomography (CT), particularly propagation-based imaging (PBI), offers superior soft tissue contrast but has historically been limited by the lack of compatible fixation techniques that preserve lung architecture post-excision. We present an adapted formaldehyde (FA) vapour fixation protocol designed to maintain human-sized lungs in a physiologically inflated and morphologically stable state. This approach prevents collapse of the delicate air\u2013tissue interfaces, a major barrier to high-fidelity phase-contrast imaging and histological correlation. Our method enables high-resolution, multiscale imaging from whole-organ PBI at 67\u00a0\u00b5m voxel size to localized subcellular synchrotron PBI at 650\u00a0nm voxel size on the same specimen, with preserved spatial relationships critical for accurate validation of imaging findings. In porcine models, FA vapour fixation maintained alveolar integrity and radiological contrast without compromising histological detail, while also avoiding the artifacts associated with liquid fixation. Crucially, the protocol allows regulation of inflation and fixation dynamics, addressing longstanding challenges in\n                    <jats:italic>ex vivo<\/jats:italic>\n                    lung imaging and enabling consistent specimen preparation across studies. This fixation technique supports biosafe stabilization of freshly explanted human lungs\u2013such as those from transplant procedures creating new opportunities for translational research on pathological tissue. By bridging high-resolution radiology and histopathology, our scalable fixation protocol establishes a standardized foundation for multimodal lung imaging and offers a critical tool for advancing both fundamental lung research and clinical diagnostics.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-23903-1","type":"journal-article","created":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T10:38:10Z","timestamp":1760956690000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Formaldehyde vapour fixation enables multiscale phase-contrast imaging and histological validation of human-sized lungs"],"prefix":"10.1038","volume":"15","author":[{"given":"Christian","family":"Dullin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johanna","family":"Reiser","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Willi L.","family":"Wagner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elena","family":"Longo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marko","family":"Pra\u0161ek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adriano","family":"Contillo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicola","family":"Sodini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diego","family":"Dreossi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Confalonieri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Salton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Confalonieri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisa","family":"Baratella","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Assunta","family":"Cova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia V.","family":"Benke","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Motiur Rahman","family":"Sagar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lorenzo","family":"D\u2019Amico","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonas","family":"Albers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angelika","family":"Svetlove","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth","family":"Duke","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatiana","family":"Flisikowska","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krzysztof","family":"Flisikowski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark O.","family":"Wielp\u00fctz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00fcrgen","family":"Biederer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans-Ulrich","family":"Kauczor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frauke","family":"Alves","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabrizio","family":"Zanconati","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuliana","family":"Tromba","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,10,20]]},"reference":[{"key":"23903_CR1","doi-asserted-by":"publisher","first-page":"6355","DOI":"10.1143\/JJAP.44.6355","volume":"44","author":"A Momose","year":"2005","unstructured":"Momose, A. 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Two lungs were obtained from an approved animal experiment using a sarcoma mini-pig model (animal permit number: 55.2-2532.Vet_02-18-33) following the completion of the study. All other lungs were sourced from a licensed slaughterhouse. These specimens underwent routine veterinary inspection and were handled under the same hygiene standards as fresh meat. Storage, transport, handling, and disposal of animal by-products were registered with the responsible veterinary authority. All procedures complied with applicable national (German) legislation. Pigs from the slaughterhouse were euthanised using a captive bolt pistol. The sarcoma model pigs received intramuscular injections of ketamine (20\u00a0mg\/kg body weight) and azaperone (2\u00a0mg\/kg body weight), were rendered unconscious by a captive bolt gun, and subsequently exsanguinated.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}},{"value":"All authors have reviewed the manuscript and gave their consent for publication.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}}],"article-number":"36475"}}