Three-dimensional Cardiomyocytes Structure Revealed By Diffusion Tensor Imaging and Its Validation Using a Tissue-Clearing Technique

Abstract

We characterized the microstructural response of the myocardium to cardiovascular disease using diffusion tensor imaging (DTI) and performed histological validation by intact, un-sectioned, three-dimensional (3D) histology using a tissue-clearing technique. The approach was validated in normal (n = 7) and ischemic (n = 8) heart failure model mice. Whole heart fiber tracking using DTI in fixed ex-vivo mouse hearts was performed, and the hearts were processed with the tissue-clearing technique. Cardiomyocytes orientation was quantified on both DTI and 3D histology. Helix angle (HA) and global HA transmurality (HAT) were calculated, and the DTI findings were confirmed with 3D histology. Global HAT was significantly reduced in the ischemic group (DTI: 0.79 ± 0.13°/% transmural depth [TD] and 3D histology: 0.84 ± 0.26°/%TD) compared with controls (DTI: 1.31 ± 0.20°/%TD and 3D histology: 1.36 ± 0.27°/%TD, all p < 0.001). On direct comparison of DTI with 3D histology for the quantitative assessment of cardiomyocytes orientation, significant correlations were observed in both per-sample (R2 = 0.803) and per-segment analyses (R2 = 0.872). We demonstrated the capability and accuracy of DTI for mapping cardiomyocytes orientation by comparison with the intact 3D histology acquired by tissue-clearing technique. DTI is a promising tool for the noninvasive characterization of cardiomyocytes architecture.

Contemporaneous 3D characterization of acute and chronic myocardial I/R injury and response

“…Applying BALANCE to our samples, tissue autofluorescence itself could be used to visualize the overall heart muscle structure allowing clear detection of cardiomyocytes and their nuclei (Fig. 2a, b) at much higher optical resolution than previously possible 16,17 .

Fig. 1Fluorophore compatible bleaching enables homogenous light sheet fluorescence microscopy (LSFM) in whole murine hearts.

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Recent Advances in Cardiac Magnetic Resonance Imaging

“…The reliability of DTI was recently validated in normal and ischemic heart failure mice models, and the myocardial fiber orientation calculated by DTI was directly compared with histology using a non-destructive 3D method with the tissue-clearing technique 57). The correlations were significant in the normal and diseased models, showing the capability and accuracy of DTI for mapping the myocardial microstructure 58). Based on these observations, the clinical application of DTI using the clinical MRI system for characterizing the myocardial fiber architecture seems possible, and it can provide additional information to conventional viability characterizations (Figure 1).…”

Aortic regurgitation in competitive athletes: The role of multimodality imaging for clinical decision-making

“…The value of newer CMR techniques such as diffusion tensor imaging in detecting myocardial architecture in VHD seems also promising. 11…”

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Figure 1.The value of multimodality imaging in differentiating volume/pressure-overload LV remodeling due to aortic valve disease from physiological adaptations to exercise; the images for diffusion tensor imaging and four-dimensional-flow aorta are reproduced from Lee et al. 11 and Ebel et al., 13 respectively.

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