Formaldehyde vapour fixation enables multiscale phase-contrast imaging and histological validation of human-sized lungs

Accurate diagnosis and characterization of lung disease increasingly rely on advanced imaging modalities capable of resolving fine microstructural details while minimizing radiation exposure. 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-tissue 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 µm voxel size to localized subcellular synchrotron PBI at 650 nm 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 ex vivo lung imaging and enabling consistent specimen preparation across studies. This fixation technique supports biosafe stabilization of freshly explanted human lungs-such 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.

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Publication Details

Journal
Scientific Reports
Published
2025-10-20
DOI
https://doi.org/10.1038/s41598-025-23903-1
Citations
1
Primary Topic
Advanced X-ray Imaging Techniques
Type
article
Field-Weighted Citation Impact
1.88
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article

Formaldehyde vapour fixation enables multiscale phase-contrast imaging and histological validation of human-sized lungs

Elisa Baratella, Lorenzo D’Amico, Angelika Svetlove, Mark Oliver Wielpütz et al.
1 citations
Scientific Reports
Advanced X-ray Imaging Techniques
1.88
article

Formaldehyde vapour fixation enables multiscale phase-contrast imaging and histological validation of human-sized lungs

Elisa Baratella, Lorenzo D’Amico, Angelika Svetlove, Mark Oliver Wielpütz, Marko Kudrna Prašek, Marco Confalonieri, Fabrizio Zanconati, Willi L. Wagner, Frauke Alves, Christian Dullin, Elena Longo, Paola Confalonieri, Elizabeth Duke, Maria Assunta Cova, Krzysztof Flisikowski, Johanna Reiser, Diego Dreossi, Francesco Salton, Nicola Sodini, Tatiana Flisikowska, Giuliana Tromba, Jonas Albers, Adriano Contillo, Claudia Benke, Hans‐Ulrich Kauczor, Md Motiur Rahman Sagar, Jürgen Biederer
article en
1 citations

Abstract

Accurate diagnosis and characterization of lung disease increasingly rely on advanced imaging modalities capable of resolving fine microstructural details while minimizing radiation exposure. 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-tissue 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 µm voxel size to localized subcellular synchrotron PBI at 650 nm 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 ex vivo lung imaging and enabling consistent specimen preparation across studies. This fixation technique supports biosafe stabilization of freshly explanted human lungs-such 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.

Scientific ReportsVol. 15(1)
Heidelberg University (DE), Universitätsmedizin Greifswald (DE), University Hospital Heidelberg (DE), Elettra-Sincrotrone Trieste S.C.p.A. (IT), Weihenstephan-Triesdorf University of Applied Sciences (DE), Christian-Albrechts-Universität zu Kiel (DE), Ospedale di Cattinara (IT), German Center for Lung Research (DE), European Molecular Biology Laboratory (DE), European Molecular Biology Laboratory (DE), Tissue Dynamics (Israel) (IL), Max Planck Institute for Multidisciplinary Sciences (DE), Technical University of Munich (DE), University of Göttingen (DE), University of Latvia (LV)
Openalex Percentile: Top 12%
Advanced X-ray Imaging Techniques
1.88
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