Circulation Journal Official Journal of the Japanese Circulation Society http://www. j-circ.or.jp ently 2 dimensional (D), whereas the heart is a 3D structure, 3D morphology could represent an additional valid source of information for ventricular non-compaction in transgenic mice. Although it is possible to use computation techniques including image registration and distortion correction to reconstruct the 3D cardiac morphology from stacked 2D sections, 26 the processes are technically demanding, laborious and time-consuming. Moreover, the 3D reconstructions still rely on 2D sections, whose destructive nature precludes their use for in vivo or in situ assessment.Non-invasive, non-destructive 3D imaging of myocardial compaction in the embryonic mouse heart is faced with two technical challenges. First, an imaging modality with the appropriate combination of tissue penetration, contrast and spatial resolution needs to be identified. The spatial resolution of magnetic resonance imaging (MRI) and computed tomography (CT) has reached microscopic scales (1 s and 10 s of he mammalian embryonic cardiac ventricles are composed of an outer layer of compact myocardium and an inner layer of trabecular myocardium. 1,2 During development, compact myocardium increases in thickness while trabeculations almost completely disappear. The persistence of a thin compact myocardium with deep intertrabecular recesses is termed ventricular non-compaction in mice, and left ventricular non-compaction (LVNC) in humans. 3-6 LVNC is a relatively common and severe genetic cardiomyopathy, 7-10 in which approximately 50% of patients show biventricular involvement. 4,11,12 Often, transgenic mice have been utilized to study LVNC. 13,14 Detailed and accurate quantification of compaction in these mice is essential to define the phenotype.Morphological analyses of the compact myocardium have been traditionally performed through histological techniques involving image analysis of cardiac sections stained with hematoxylin and eosin (H&E). Background: Ventricular non-compaction is characterized by a thin layer of compact ventricular myocardium and it is an important abnormality in the mouse heart. It is reminiscent of left ventricular non-compaction, a fairly common human congenital cardiomyopathy. Non-compaction in transgenic mice has been classically evaluated by measuring the thickness of the compact myocardium through histological techniques involving image analysis of 2-dimensional (D) sections. Given the 3D nature of the heart, the aim of this study was to determine whether a technique for the non-destructive, 3D assessment of the mouse embryonic compact myocardium could be developed.