Abstract

Micro-CT scan images enhanced by iodine staining provide high-resolution visualisation of soft tissues in laboratory mice. We have compared Micro-CT scan-derived left ventricular (LV) mass with dissection and weighing. Ex-vivo micro-CT scan images of the mouse hearts were obtained following staining by iodine. The LV was segmented and its volume was assessed using a semi-automated method by Drishti software. The left ventricle was then dissected in the laboratory and its actual weight was measured and compared against the estimated results. LV mass was calculated multiplying its estimated volume and myocardial specific gravity. Thirty-five iodine-stained post-natal mouse hearts were studied. Mice were of either sex and 68 to 352 days old (median age 202 days with interquartile range 103 to 245 days) at the time of sacrifice. Samples were from 20 genetically diverse strains. Median mouse body weight was 29 g with interquartile range 24 to 34 g. Left Ventricular weights ranged from 40.0 to 116.7 mg. The segmented LV mass estimated from micro-CT scan and directly measured dissected LV mass were strongly correlated (R2 = 0. 97). Segmented LV mass derived from Micro-CT images was very similar to the physically dissected LV mass (mean difference = 0.09 mg; 95% confidence interval − 3.29 mg to 3.1 mg). Micro-CT scanning provides a non-destructive, efficient and accurate visualisation tool for anatomical analysis of animal heart models of human cardiovascular conditions. Iodine-stained soft tissue imaging empowers researchers to perform qualitative and quantitative assessment of the cardiac structures with preservation of the samples for future histological analysis.

Highlights

  • Micro-computerised tomography scan (CT scan) images enhanced by iodine staining provide high-resolution visualisation of soft tissues in laboratory mice

  • Estimation of left ventricular (LV) mass can be performed by several imaging modalities including echocardiography, cardiac computerised tomography scan (CT scan) and cardiac magnetic resonance (CMR)[4]

  • Our aim was to develop a non-destructive method of estimating ex-vivo left ventricular mass/tissue-volume in mice as part of a project aimed at identifying genes that contribute to left ventricular hypertrophy in the Collaborative Cross (CC)[12]

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Summary

Introduction

Micro-CT scan images enhanced by iodine staining provide high-resolution visualisation of soft tissues in laboratory mice. Iodine-stained soft tissue imaging empowers researchers to perform qualitative and quantitative assessment of the cardiac structures with preservation of the samples for future histological analysis. The initial breeding s­ ites[13] were in Oak Ridge (USA), Perth (Australia) and Nairobi (Kenya) and these populations have moved to Chapel H­ ill[13] (USA), Beijing (China) and Tel A­ viv[13] (Israel) respectively This has created some difficulty with access to the strains for phenotyping. We applied staining enhanced micro-CT scan for high resolution imaging of cardiovascular structure in laboratory mice to reconstruct the LV and measure the LV tissue–volume/mass. Virtual dissection of micro-CT 3-dimensional images provides non-destructive measurement of left-ventricular tissue volume allowing both secondary uses and repeated measurements neither of which are possible with physical dissection. Our results support the use of micro-CT to estimate LV tissue volume, in circumstances where it is desirable to leave the preserved specimen intact for other subsequent use

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