Abstract
We present an analysis of imaging murine embryos at various embryonic developmental stages (embryonic day 9.5, 11.5, and 13.5) by optical coherence tomography (OCT) and optical projection tomography (OPT). We demonstrate that while OCT was capable of rapid high-resolution live 3D imaging, its limited penetration depth prevented visualization of deeper structures, particularly in later stage embryos. In contrast, OPT was able to image the whole embryos, but could not be used in vivo because the embryos must be fixed and cleared. Moreover, the fixation process significantly altered the embryo morphology, which was quantified by the volume of the eye-globes before and after fixation. All of these factors should be weighed when determining which imaging modality one should use to achieve particular goals of a study.
Highlights
Murine models have provided enormous insights into the mechanisms that underlie mammalian development and birth defects
Previous investigations have successfully utilized Optical coherence tomography (OCT) to study cardiac [20, 58], ocular [50, 54], vascular [36], and neural [59] development in murine embryos as well as visualize microinjections of drugs [60]. Additional methods such as rotational imaging could be combined with OCT to allow multi-angle embryonic imaging [29]
Optical projection tomography (OPT) allows for acquisition of high resolution full body images of murine embryos with excellent spatial resolution and contrast and with minimal shadowing artifacts due to back-projection reconstruction after multi-angle projection acquisition
Summary
Murine models have provided enormous insights into the mechanisms that underlie mammalian development and birth defects. The development and adaptation of imaging techniques such as ultrasound biomicroscopy (UBM) [5, 6], micro X-ray computed tomography (micro-CT) [6, 7], and micro magnetic resonance imaging (micro-MRI) [8, 9], has proven invaluable at providing phenotypic information of small mammalian embryos noninvasively. In utero micro-MRI has been performed at the cost of spatial resolution, and the gating and registration methods required to remove maternal motion resulted in extended acquisition times of ~2 hours [11]
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