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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

Visualization of the aortic sinus.(A) In a slice stained with Movat’s pentachrome, the valves and several atherosclerotic plaques of different size (arrows) can be observed. Here, blue-green stains for ground substance (non-cellular components of the extracellular matrix), blue-black stains for nuclei and elastic fibers, red stains for muscle, intense red stains for fibrinoid material and fibrin and green-yellow stains for collagen and reticulin fibers. (B) A DPC image at the same location that was taken from a 3D dataset allows the identification of the same structures, with sufficient contrast between the background, valves, plaques and vessel walls. (C) A slice from the 3D dataset with a slightly different orientation allows for an improved overview of the plaque. (D,E) Plaques cannot be observed in the Movat’s pentachrome staining or DPC image of the aortic sinus of a WT control mouse. Scale bars indicate 200 μm.
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f2: Visualization of the aortic sinus.(A) In a slice stained with Movat’s pentachrome, the valves and several atherosclerotic plaques of different size (arrows) can be observed. Here, blue-green stains for ground substance (non-cellular components of the extracellular matrix), blue-black stains for nuclei and elastic fibers, red stains for muscle, intense red stains for fibrinoid material and fibrin and green-yellow stains for collagen and reticulin fibers. (B) A DPC image at the same location that was taken from a 3D dataset allows the identification of the same structures, with sufficient contrast between the background, valves, plaques and vessel walls. (C) A slice from the 3D dataset with a slightly different orientation allows for an improved overview of the plaque. (D,E) Plaques cannot be observed in the Movat’s pentachrome staining or DPC image of the aortic sinus of a WT control mouse. Scale bars indicate 200 μm.

Mentions: Movat’s pentachrome staining of histological cross-sections of the aortic sinus showed intermediate-stage atherosclerotic plaques, defined by the presence of necrotic cores containing cholesterol clefts and thin fibrous caps. The DPC images at the same location matched well with these histological slices (Fig. 2). The vessel wall, plaques and valves were readily observed, while the thin caps of the intermediate plaques were difficult to observe 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)

Visualization of the aortic sinus.(A) In a slice stained with Movat’s pentachrome, the valves and several atherosclerotic plaques of different size (arrows) can be observed. Here, blue-green stains for ground substance (non-cellular components of the extracellular matrix), blue-black stains for nuclei and elastic fibers, red stains for muscle, intense red stains for fibrinoid material and fibrin and green-yellow stains for collagen and reticulin fibers. (B) A DPC image at the same location that was taken from a 3D dataset allows the identification of the same structures, with sufficient contrast between the background, valves, plaques and vessel walls. (C) A slice from the 3D dataset with a slightly different orientation allows for an improved overview of the plaque. (D,E) Plaques cannot be observed in the Movat’s pentachrome staining or DPC image of the aortic sinus of a WT control mouse. Scale bars indicate 200 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Visualization of the aortic sinus.(A) In a slice stained with Movat’s pentachrome, the valves and several atherosclerotic plaques of different size (arrows) can be observed. Here, blue-green stains for ground substance (non-cellular components of the extracellular matrix), blue-black stains for nuclei and elastic fibers, red stains for muscle, intense red stains for fibrinoid material and fibrin and green-yellow stains for collagen and reticulin fibers. (B) A DPC image at the same location that was taken from a 3D dataset allows the identification of the same structures, with sufficient contrast between the background, valves, plaques and vessel walls. (C) A slice from the 3D dataset with a slightly different orientation allows for an improved overview of the plaque. (D,E) Plaques cannot be observed in the Movat’s pentachrome staining or DPC image of the aortic sinus of a WT control mouse. Scale bars indicate 200 μm.
Mentions: Movat’s pentachrome staining of histological cross-sections of the aortic sinus showed intermediate-stage atherosclerotic plaques, defined by the presence of necrotic cores containing cholesterol clefts and thin fibrous caps. The DPC images at the same location matched well with these histological slices (Fig. 2). The vessel wall, plaques and valves were readily observed, while the thin caps of the intermediate plaques were difficult to observe 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