Cryogenic contrast-enhanced microCT enables nondestructive 3D quantitative histopathology of soft biological tissues

Arne Maes, Camille Pestiaux, Alice Marino, Tim Balcaen, Lisa Leyssens, Sarah Vangrunderbeeck, Grzegorz Pyka, Wim M. De Borggraeve, Luc Bertrand, Christophe Beauloye, Sandrine Horman, Martine Wevers and Greet Kerckhofs ()
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Arne Maes: KU Leuven
Camille Pestiaux: UCLouvain
Alice Marino: UCLouvain
Tim Balcaen: UCLouvain
Lisa Leyssens: UCLouvain
Sarah Vangrunderbeeck: UCLouvain
Grzegorz Pyka: UCLouvain
Wim M. De Borggraeve: KU Leuven
Luc Bertrand: UCLouvain
Christophe Beauloye: University Hospital Saint-Luc
Sandrine Horman: UCLouvain
Martine Wevers: KU Leuven
Greet Kerckhofs: KU Leuven

Nature Communications, 2022, vol. 13, issue 1, 1-14

Abstract: Abstract Biological tissues comprise a spatially complex structure, composition and organization at the microscale, named the microstructure. Given the close structure-function relationships in tissues, structural characterization is essential to fully understand the functioning of healthy and pathological tissues, as well as the impact of possible treatments. Here, we present a nondestructive imaging approach to perform quantitative 3D histo(patho)logy of biological tissues, termed Cryogenic Contrast-Enhanced MicroCT (cryo-CECT). By combining sample staining, using an X-ray contrast-enhancing staining agent, with freezing the sample at the optimal freezing rate, cryo-CECT enables 3D visualization and structural analysis of individual tissue constituents, such as muscle and collagen fibers. We applied cryo-CECT on murine hearts subjected to pressure overload following transverse aortic constriction surgery. Cryo-CECT allowed to analyze, in an unprecedented manner, the orientation and diameter of the individual muscle fibers in the entire heart, as well as the 3D localization of fibrotic regions within the myocardial layers. We foresee further applications of cryo-CECT in the optimization of tissue/food preservation and donor banking, showing that cryo-CECT also has clinical and industrial potential.

Date: 2022
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DOI: 10.1038/s41467-022-34048-4

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