Non-destructive 3D characterization of the blood vessel wall microstructure in different species and blood vessel types using contrast-enhanced microCT and comparison with synthetic vascular grafts

Leyssens, Lisa; Balcaen, Tim; Pétréa, Maïté; Ayllón, Natalia Béjar; Aazmani, Walid El; Pierpont, Alix de; Pyka, Grzegorz; Lacroix, Valérie; Kerckhofs, Greet

doi:10.1016/j.actbio.2023.04.013

Cited by 8 publications

(2 citation statements)

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“…Given the substantial tissue shrinkage during sample preparation for classical 2D histology (Supplementary Figure 10 ), a quantitative comparison between cryo-CECT and classical 2D histology could lead to erroneous conclusions. Similar observations regarding tissue shrinkage due to paraffin embedding have been made for murine hearts [ 28 , 38 ] and for porcine aortic wall tissue [ 52 ]. Also, the current semiautomatic segmentation method was not able to keep track of the tubule when it transitioned from thick descending limb to thin descending limb in the inner medulla.…”

Section: Discussionsupporting

confidence: 71%

“…For instance, GATA3 and podoplanin immunohistochemical staining was still successful on kidneys that underwent cryo-CECT (Supplementary Figure 9 ). In addition, several (immuno)histochemical stainings following (cryo-)CECT based on Hf-WD POM have been described in literature, including CD31 immunostain [ 28 , 51 ], Verhoeff's Van Gieson stain [ 52 ], Sirius red stain [ 28 ], and Masson's trichrome [ 28 ]. Now that the proof of concept has been established, other murine kidney disease models with more severe/localized morphologic changes, such as IRI, sepsis-induced, or autoimmune-associated AKI, could be studied using the cryo-CECT technique.…”

Section: Discussionmentioning

confidence: 99%

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X-Ray-Based 3D Histopathology of the Kidney Using Cryogenic Contrast-Enhanced MicroCT

Maes,

Borgel,

Braconnier

et al. 2024

International Journal of Biomedical Imaging

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The kidney’s microstructure, which comprises a highly convoluted tubular and vascular network, can only be partially revealed using classical 2D histology. Considering that the kidney’s microstructure is closely related to its function and is often affected by pathologies, there is a need for powerful and high-resolution 3D imaging techniques to visualize the microstructure. Here, we present how cryogenic contrast-enhanced microCT (cryo-CECT) allowed 3D visualization of glomeruli, tubuli, and vasculature. By comparing different contrast-enhancing staining agents and freezing protocols, we found that the preferred sample preparation protocol was the combination of staining with 1:2 hafnium(IV)-substituted Wells-Dawson polyoxometalate and freezing by submersion in isopentane at −78°C. This optimized protocol showed to be highly sensitive, allowing to detect small pathology-induced microstructural changes in a mouse model of mild trauma-related acute kidney injury after thorax trauma and hemorrhagic shock. In summary, we demonstrated that cryo-CECT is an effective 3D histopathological tool that allows to enhance our understanding of kidney tissue microstructure and their related function.

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Section: Discussionsupporting

confidence: 71%