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Quantitative 3-dimensional imaging of murine neointimal and atherosclerotic lesions by optical projection tomography.

Kirkby NS, Low L, Seckl JR, Walker BR, Webb DJ, Hadoke PW - PLoS ONE (2011)

Bottom Line: Planimetric measurements of lesion area correlated well with those made from histological sections subsequently produced from the same vessels (wire-injury: R²  =  0.92; ligation-injury: R²  =  0.89; atherosclerosis: R²  =  0.85), confirming both the accuracy of this methodology and its non-destructive nature.It was also possible to record volumetric measurements of lesion and lumen and these were highly reproducible between scans (coefficient of variation  =  5.36%, 11.39% and 4.79% for wire- and ligation-injury and atherosclerosis, respectively).These data demonstrate the eminent suitability of OPT for imaging of atherosclerotic and neointimal lesion formation, providing a much needed means for the routine 3-dimensional analysis of vascular morphology in studies of this type.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular Science, University of Edinburgh, Edinburgh, United Kingdom.

ABSTRACT

Objective: Traditional methods for the analysis of vascular lesion formation are labour intensive to perform - restricting study to 'snapshots' within each vessel. This study was undertaken to determine the suitability of optical projection tomographic (OPT) imaging for the 3-dimensional representation and quantification of intimal lesions in mouse arteries.

Methods and results: Vascular injury was induced by wire-insertion or ligation of the mouse femoral artery or administration of an atherogenic diet to apoE-deficient mice. Lesion formation was examined by OPT imaging of autofluorescent emission. Lesions could be clearly identified and distinguished from the underlying vascular wall. Planimetric measurements of lesion area correlated well with those made from histological sections subsequently produced from the same vessels (wire-injury: R²  =  0.92; ligation-injury: R²  =  0.89; atherosclerosis: R²  =  0.85), confirming both the accuracy of this methodology and its non-destructive nature. It was also possible to record volumetric measurements of lesion and lumen and these were highly reproducible between scans (coefficient of variation  =  5.36%, 11.39% and 4.79% for wire- and ligation-injury and atherosclerosis, respectively).

Conclusions: These data demonstrate the eminent suitability of OPT for imaging of atherosclerotic and neointimal lesion formation, providing a much needed means for the routine 3-dimensional analysis of vascular morphology in studies of this type.

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Comparison of planimetric measurements of lesion size between OPT reconstructions and histological sections.Planimetric measurements of lesion size recorded from OPT and histology image sets correlate strongly for all lesion types; Bland-Altman analysis indicates that this is unbiased for wire- (a) and ligation-injured femoral arteries (b) and that OPT measurements have a consistent positive bias in atherosclerotic brachiocephalic arteries (c).
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pone-0016906-g003: Comparison of planimetric measurements of lesion size between OPT reconstructions and histological sections.Planimetric measurements of lesion size recorded from OPT and histology image sets correlate strongly for all lesion types; Bland-Altman analysis indicates that this is unbiased for wire- (a) and ligation-injured femoral arteries (b) and that OPT measurements have a consistent positive bias in atherosclerotic brachiocephalic arteries (c).

Mentions: To validate the interpretation of OPT reconstructions following tomographic imaging, the same arteries were processed for histological examination. For both neointimal (Figure 1c) and atherosclerotic lesions (Figure 2c), histological sections and corresponding OPT reconstructions were strikingly similar. We compared planimetric measurements of lesion area obtained by each method. For wire- and ligation-induced neointimal lesions and atherosclerotic plaques, these correlated strongly by linear regression (R2 = 0.92, R2 = 0.89 and R2 = 0.85, respectively) and the slope of these relationships did not differ from 1 (p = 0.49, p = 0.58 and p = 0.88, respectively). Bland-Altman analysis indicated no obvious bias in measurements of neointimal lesions induced by either injury (Figure 3a,b) but a small positive bias in OPT-derived measurements of lesion size in the brachiocephalic artery (Figure 3c), possibly reflecting shrinkage of these larger vessels during histological processing. Importantly, however, this bias was independent of lesion size indicating that measurements made in this way are comparable within and between vessels. We also confirmed that the OPT imaging procedure did not compromise subsequent analysis of lesion composition. In vessels previously subjected to OPT scanning, (immuno)staining for markers of smooth muscle (α-smooth muscle actin), macrophages (Mac2) and collagen (picro-sirius red) all occurred with the expected intensity and distribution (Figures 2c).


Quantitative 3-dimensional imaging of murine neointimal and atherosclerotic lesions by optical projection tomography.

Kirkby NS, Low L, Seckl JR, Walker BR, Webb DJ, Hadoke PW - PLoS ONE (2011)

Comparison of planimetric measurements of lesion size between OPT reconstructions and histological sections.Planimetric measurements of lesion size recorded from OPT and histology image sets correlate strongly for all lesion types; Bland-Altman analysis indicates that this is unbiased for wire- (a) and ligation-injured femoral arteries (b) and that OPT measurements have a consistent positive bias in atherosclerotic brachiocephalic arteries (c).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3040742&req=5

pone-0016906-g003: Comparison of planimetric measurements of lesion size between OPT reconstructions and histological sections.Planimetric measurements of lesion size recorded from OPT and histology image sets correlate strongly for all lesion types; Bland-Altman analysis indicates that this is unbiased for wire- (a) and ligation-injured femoral arteries (b) and that OPT measurements have a consistent positive bias in atherosclerotic brachiocephalic arteries (c).
Mentions: To validate the interpretation of OPT reconstructions following tomographic imaging, the same arteries were processed for histological examination. For both neointimal (Figure 1c) and atherosclerotic lesions (Figure 2c), histological sections and corresponding OPT reconstructions were strikingly similar. We compared planimetric measurements of lesion area obtained by each method. For wire- and ligation-induced neointimal lesions and atherosclerotic plaques, these correlated strongly by linear regression (R2 = 0.92, R2 = 0.89 and R2 = 0.85, respectively) and the slope of these relationships did not differ from 1 (p = 0.49, p = 0.58 and p = 0.88, respectively). Bland-Altman analysis indicated no obvious bias in measurements of neointimal lesions induced by either injury (Figure 3a,b) but a small positive bias in OPT-derived measurements of lesion size in the brachiocephalic artery (Figure 3c), possibly reflecting shrinkage of these larger vessels during histological processing. Importantly, however, this bias was independent of lesion size indicating that measurements made in this way are comparable within and between vessels. We also confirmed that the OPT imaging procedure did not compromise subsequent analysis of lesion composition. In vessels previously subjected to OPT scanning, (immuno)staining for markers of smooth muscle (α-smooth muscle actin), macrophages (Mac2) and collagen (picro-sirius red) all occurred with the expected intensity and distribution (Figures 2c).

Bottom Line: Planimetric measurements of lesion area correlated well with those made from histological sections subsequently produced from the same vessels (wire-injury: R²  =  0.92; ligation-injury: R²  =  0.89; atherosclerosis: R²  =  0.85), confirming both the accuracy of this methodology and its non-destructive nature.It was also possible to record volumetric measurements of lesion and lumen and these were highly reproducible between scans (coefficient of variation  =  5.36%, 11.39% and 4.79% for wire- and ligation-injury and atherosclerosis, respectively).These data demonstrate the eminent suitability of OPT for imaging of atherosclerotic and neointimal lesion formation, providing a much needed means for the routine 3-dimensional analysis of vascular morphology in studies of this type.

View Article: PubMed Central - PubMed

Affiliation: Centre for Cardiovascular Science, University of Edinburgh, Edinburgh, United Kingdom.

ABSTRACT

Objective: Traditional methods for the analysis of vascular lesion formation are labour intensive to perform - restricting study to 'snapshots' within each vessel. This study was undertaken to determine the suitability of optical projection tomographic (OPT) imaging for the 3-dimensional representation and quantification of intimal lesions in mouse arteries.

Methods and results: Vascular injury was induced by wire-insertion or ligation of the mouse femoral artery or administration of an atherogenic diet to apoE-deficient mice. Lesion formation was examined by OPT imaging of autofluorescent emission. Lesions could be clearly identified and distinguished from the underlying vascular wall. Planimetric measurements of lesion area correlated well with those made from histological sections subsequently produced from the same vessels (wire-injury: R²  =  0.92; ligation-injury: R²  =  0.89; atherosclerosis: R²  =  0.85), confirming both the accuracy of this methodology and its non-destructive nature. It was also possible to record volumetric measurements of lesion and lumen and these were highly reproducible between scans (coefficient of variation  =  5.36%, 11.39% and 4.79% for wire- and ligation-injury and atherosclerosis, respectively).

Conclusions: These data demonstrate the eminent suitability of OPT for imaging of atherosclerotic and neointimal lesion formation, providing a much needed means for the routine 3-dimensional analysis of vascular morphology in studies of this type.

Show MeSH
Related in: MedlinePlus