4D optical coherence tomography of the embryonic heart using gated imaging

Computed tomography (CT), ultrasound, and magnetic resonance imaging have been used to image and diagnose diseases of the human heart. By gating the acquisition of the images to the heart cycle (gated imaging), these modalities enable one to produce 3D images of the heart without significant motion artifact and to more accurately calculate various parameters such as ejection fractions [1-3]. Unfortunately, these imaging modalities give inadequate resolution when investigating embryonic development in animal models. Defects in developmental mechanisms during embryogenesis have long been thought to result in congenital cardiac anomalies. Our understanding of normal mechanisms of heart development and how abnormalities can lead to defects has been hampered by our inability to detect anatomic and physiologic changes in these small (<2mm) organs. Optical coherence tomography (OCT) has made it possible to visualize internal structures of the living embryonic heart with high-resolution in two- and threedimensions. OCT offers higher resolution than ultrasound (30 um axial, 90 um lateral) and magnetic resonance microscopy (25 um axial, 31 um lateral) [4, 5], with greater depth penetration over confocal microscopy (200 um). Optical coherence tomography (OCT) uses back reflected light from a sample to create an image with axial resolutions ranging from 2-15 um, while penetrating 1-2 mm in depth [6]. In the past, OCT groups estimated ejection fractions using 2D images in a Xenopus laevis [7], created 3D renderings of chick embryo hearts [8], and used a gated reconstruction technique to produce 2D Doppler OCT image of an in vivo Xenopus laevis heart [9]. In this paper we present a gated imaging system that allowed us to produce a 16-frame 3D movie of a beating chick embryo heart. The heart was excised from a day two (stage 13) chicken embryo and electrically paced at 1 Hz. We acquired 2D images (B-scans) in 62.5 ms, which provides enough temporal resolution to distinguish end-contraction from end-relaxation. After acquiring the image set, we were able to measure the ejection fraction.