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Ultra-high-resolution 3D imaging of atherosclerosis in mice with synchrotron differential phase contrast: a proof of concept study.

Bonanno G, Coppo S, Modregger P, Pellegrin M, Stuber A, Stampanoni M, Mazzolai L, Stuber M, van Heeswijk RB - Sci Rep (2015)

Bottom Line: The DPC imaging allowed for the visualization of complex structures such as the coronary arteries and their branches, the thin fibrous cap of atherosclerotic plaques as well as the chordae tendineae.The coronary vessel wall thickness ranged from 37.4 ± 5.6 μm in proximal coronary arteries to 13.6 ± 3.3 μm in distal branches.No consistent differences in coronary vessel wall thickness were detected between the wild-type and atherosclerotic hearts in this proof-of-concept study, although the standard deviation in the atherosclerotic mice was higher in most segments, consistent with the observation of occasional focal vessel wall thickening.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Radiology, University Hospital (CHUV) and University (UNIL) of Lausanne, Switzerland [2] Center for Biomedical Imaging (CIBM), Lausanne, Switzerland.

ABSTRACT
The goal of this study was to investigate the performance of 3D synchrotron differential phase contrast (DPC) imaging for the visualization of both macroscopic and microscopic aspects of atherosclerosis in the mouse vasculature ex vivo. The hearts and aortas of 2 atherosclerotic and 2 wild-type control mice were scanned with DPC imaging with an isotropic resolution of 15 μm. The coronary artery vessel walls were segmented in the DPC datasets to assess their thickness, and histological staining was performed at the level of atherosclerotic plaques. The DPC imaging allowed for the visualization of complex structures such as the coronary arteries and their branches, the thin fibrous cap of atherosclerotic plaques as well as the chordae tendineae. The coronary vessel wall thickness ranged from 37.4 ± 5.6 μm in proximal coronary arteries to 13.6 ± 3.3 μm in distal branches. No consistent differences in coronary vessel wall thickness were detected between the wild-type and atherosclerotic hearts in this proof-of-concept study, although the standard deviation in the atherosclerotic mice was higher in most segments, consistent with the observation of occasional focal vessel wall thickening. Overall, DPC imaging of the cardiovascular system of the mice allowed for a simultaneous detailed 3D morphological assessment of both large structures and microscopic details.

No MeSH data available.


Related in: MedlinePlus

Overview of the DPC imaging and histology in the mouse aorta and branches.(A,B) En face photos of the excised aortas and branches of a WT control and ApoE−/− mouse. In the latter, plaques can be clearly observed in the inner curvature of the aorta and the brachiocephalic artery (arrows). (C) DPC image of the same mouse aorta as in B). A large plaque that obstructs most of the brachiocephalic artery (solid arrow) and small plaques on the inner curvature (dotted arrow) can be clearly observed. (D) Longitudinal view of a 3D segmentation of the plaque within the brachiocephalic artery; the volume of the plaque (yellow) was calculated to be 209 nL. (E) Movat’s pentachrome staining of the same large brachiocephalic plaque. While the plaque obstructs most of the artery and contains a large necrotic core with numerous cholesterol clefts, its overlying cap (arrows) is fairly thick. (F) Zoom-in of the DPC image at the same location, where the cap with a thickness of 28.7 ± 8.7 μm can also be observed (arrows). Although different intensities can be observed inside the plaque, the exact makeup is harder to discern.
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f3: Overview of the DPC imaging and histology in the mouse aorta and branches.(A,B) En face photos of the excised aortas and branches of a WT control and ApoE−/− mouse. In the latter, plaques can be clearly observed in the inner curvature of the aorta and the brachiocephalic artery (arrows). (C) DPC image of the same mouse aorta as in B). A large plaque that obstructs most of the brachiocephalic artery (solid arrow) and small plaques on the inner curvature (dotted arrow) can be clearly observed. (D) Longitudinal view of a 3D segmentation of the plaque within the brachiocephalic artery; the volume of the plaque (yellow) was calculated to be 209 nL. (E) Movat’s pentachrome staining of the same large brachiocephalic plaque. While the plaque obstructs most of the artery and contains a large necrotic core with numerous cholesterol clefts, its overlying cap (arrows) is fairly thick. (F) Zoom-in of the DPC image at the same location, where the cap with a thickness of 28.7 ± 8.7 μm can also be observed (arrows). Although different intensities can be observed inside the plaque, the exact makeup is harder to discern.

Mentions: DPC images of the aortic arches and their branches also matched well with the en face photos (Fig. 3). Atherosclerotic plaques with different sizes could be recognized (Fig. 3C) and segmented (Fig. 3D). In large plaques, the fibrous cap could be discerned from the necrotic core, and individual layers of smooth muscle cells in the media could be differentiated (Fig. 3E,F). However, individual cholesterol crystals that were visible in the histological sections could not readily be appreciated in the DPC images.


Ultra-high-resolution 3D imaging of atherosclerosis in mice with synchrotron differential phase contrast: a proof of concept study.

Bonanno G, Coppo S, Modregger P, Pellegrin M, Stuber A, Stampanoni M, Mazzolai L, Stuber M, van Heeswijk RB - Sci Rep (2015)

Overview of the DPC imaging and histology in the mouse aorta and branches.(A,B) En face photos of the excised aortas and branches of a WT control and ApoE−/− mouse. In the latter, plaques can be clearly observed in the inner curvature of the aorta and the brachiocephalic artery (arrows). (C) DPC image of the same mouse aorta as in B). A large plaque that obstructs most of the brachiocephalic artery (solid arrow) and small plaques on the inner curvature (dotted arrow) can be clearly observed. (D) Longitudinal view of a 3D segmentation of the plaque within the brachiocephalic artery; the volume of the plaque (yellow) was calculated to be 209 nL. (E) Movat’s pentachrome staining of the same large brachiocephalic plaque. While the plaque obstructs most of the artery and contains a large necrotic core with numerous cholesterol clefts, its overlying cap (arrows) is fairly thick. (F) Zoom-in of the DPC image at the same location, where the cap with a thickness of 28.7 ± 8.7 μm can also be observed (arrows). Although different intensities can be observed inside the plaque, the exact makeup is harder to discern.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499839&req=5

f3: Overview of the DPC imaging and histology in the mouse aorta and branches.(A,B) En face photos of the excised aortas and branches of a WT control and ApoE−/− mouse. In the latter, plaques can be clearly observed in the inner curvature of the aorta and the brachiocephalic artery (arrows). (C) DPC image of the same mouse aorta as in B). A large plaque that obstructs most of the brachiocephalic artery (solid arrow) and small plaques on the inner curvature (dotted arrow) can be clearly observed. (D) Longitudinal view of a 3D segmentation of the plaque within the brachiocephalic artery; the volume of the plaque (yellow) was calculated to be 209 nL. (E) Movat’s pentachrome staining of the same large brachiocephalic plaque. While the plaque obstructs most of the artery and contains a large necrotic core with numerous cholesterol clefts, its overlying cap (arrows) is fairly thick. (F) Zoom-in of the DPC image at the same location, where the cap with a thickness of 28.7 ± 8.7 μm can also be observed (arrows). Although different intensities can be observed inside the plaque, the exact makeup is harder to discern.
Mentions: DPC images of the aortic arches and their branches also matched well with the en face photos (Fig. 3). Atherosclerotic plaques with different sizes could be recognized (Fig. 3C) and segmented (Fig. 3D). In large plaques, the fibrous cap could be discerned from the necrotic core, and individual layers of smooth muscle cells in the media could be differentiated (Fig. 3E,F). However, individual cholesterol crystals that were visible in the histological sections could not readily be appreciated in the DPC images.

Bottom Line: The DPC imaging allowed for the visualization of complex structures such as the coronary arteries and their branches, the thin fibrous cap of atherosclerotic plaques as well as the chordae tendineae.The coronary vessel wall thickness ranged from 37.4 ± 5.6 μm in proximal coronary arteries to 13.6 ± 3.3 μm in distal branches.No consistent differences in coronary vessel wall thickness were detected between the wild-type and atherosclerotic hearts in this proof-of-concept study, although the standard deviation in the atherosclerotic mice was higher in most segments, consistent with the observation of occasional focal vessel wall thickening.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Radiology, University Hospital (CHUV) and University (UNIL) of Lausanne, Switzerland [2] Center for Biomedical Imaging (CIBM), Lausanne, Switzerland.

ABSTRACT
The goal of this study was to investigate the performance of 3D synchrotron differential phase contrast (DPC) imaging for the visualization of both macroscopic and microscopic aspects of atherosclerosis in the mouse vasculature ex vivo. The hearts and aortas of 2 atherosclerotic and 2 wild-type control mice were scanned with DPC imaging with an isotropic resolution of 15 μm. The coronary artery vessel walls were segmented in the DPC datasets to assess their thickness, and histological staining was performed at the level of atherosclerotic plaques. The DPC imaging allowed for the visualization of complex structures such as the coronary arteries and their branches, the thin fibrous cap of atherosclerotic plaques as well as the chordae tendineae. The coronary vessel wall thickness ranged from 37.4 ± 5.6 μm in proximal coronary arteries to 13.6 ± 3.3 μm in distal branches. No consistent differences in coronary vessel wall thickness were detected between the wild-type and atherosclerotic hearts in this proof-of-concept study, although the standard deviation in the atherosclerotic mice was higher in most segments, consistent with the observation of occasional focal vessel wall thickening. Overall, DPC imaging of the cardiovascular system of the mice allowed for a simultaneous detailed 3D morphological assessment of both large structures and microscopic details.

No MeSH data available.


Related in: MedlinePlus