Article

Optical Coherence Tomography for live imaging of mammalian development

Department of Molecular Physiology and Biophysics, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, United States.
Current opinion in genetics & development (Impact Factor: 8.57). 09/2011; 21(5):579-84. DOI: 10.1016/j.gde.2011.09.004
Source: PubMed

ABSTRACT Understanding the nature and mechanism of congenital defects of the different organ systems in humans has heavily relied on the analysis of the corresponding mutant phenotypes in rodent models. Optical Coherence Tomography (OCT) has recently emerged as a powerful tool to study early embryonic development. This non-invasive optical methodology does not require labeling and allows visualization of embryonic tissues with single cell resolution. Here, we will discuss how OCT can be applied for structural imaging of early mouse and rat embryos in static culture, cardiodynamic and blood flow analysis, and in utero embryonic imaging at later stages of gestation, demonstrating how OCT can be used to assess structural and functional birth defects in mammalian models.

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Available from: Mary E Dickinson, Mar 27, 2014
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    • "Light sheet fluorescence microscopy (LSFM) or selective plane illumination microscopy (SPIM) employs a focused light sheet of excitation light which enables fast and high resolution imaging of small samples with minimal photobleaching; perfect for time lapse 3D images of development of live embryos [12]. Optical Coherence Tomography (OCT) is an interferometric technique to measure the optical scattering of tissue that can acquire high resolution (2–10 µm) 3D data sets at suitable depths (1–3 mm) for ex-vivo and in-vivo imaging of embryos early in development [13]–[15]. "
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    ABSTRACT: Optical projection tomography (OPT) is an imaging modality that has, in the last decade, answered numerous biological questions owing to its ability to view gene expression in 3 dimensions (3D) at high resolution for samples up to several cm(3). This has increased demand for a cabinet OPT system, especially for mouse embryo phenotyping, for which OPT was primarily designed for. The Medical Research Council (MRC) Technology group (UK) released a commercial OPT system, constructed by Skyscan, called the Bioptonics OPT 3001 scanner that was installed in a limited number of locations. The Bioptonics system has been discontinued and currently there is no commercial OPT system available. Therefore, a few research institutions have built their own OPT system, choosing parts and a design specific to their biological applications. Some of these custom built OPT systems are preferred over the commercial Bioptonics system, as they provide improved performance based on stable translation and rotation stages and up to date CCD cameras coupled with objective lenses of high numerical aperture, increasing the resolution of the images. Here, we present a detailed description of a custom built OPT system that is robust and easy to build and install. Included is a hardware parts list, instructions for assembly, a description of the acquisition software and a free download site, and methods for calibration. The described OPT system can acquire a full 3D data set in 10 minutes at 6.7 micron isotropic resolution. The presented guide will hopefully increase adoption of OPT throughout the research community, for the OPT system described can be implemented by personnel with minimal expertise in optics or engineering who have access to a machine shop.
    PLoS ONE 09/2013; 8(9):e73491. DOI:10.1371/journal.pone.0073491 · 3.23 Impact Factor
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    • "magnetic resonance imaging (MRI) (Schneider et al., 2004; Dazai et al., 2011)] and others that are on the cutting edge of dynamic embryo imaging [e.g. optical coherence tomography (OCT) (Larina et al., 2011)]. When discussing the relative merits of a platform, an obvious driver is that whole-embryo coverage is obtained to ensure all organ systems are observed. "
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    ABSTRACT: Identifying genes that are important for embryo development is a crucial first step towards understanding their many functions in driving the ordered growth, differentiation and organogenesis of embryos. It can also shed light on the origins of developmental disease and congenital abnormalities. Current international efforts to examine gene function in the mouse provide a unique opportunity to pinpoint genes that are involved in embryogenesis, owing to the emergence of embryonic lethal knockout mutants. Through internationally coordinated efforts, the International Knockout Mouse Consortium (IKMC) has generated a public resource of mouse knockout strains and, in April 2012, the International Mouse Phenotyping Consortium (IMPC), supported by the EU InfraCoMP programme, convened a workshop to discuss developing a phenotyping pipeline for the investigation of embryonic lethal knockout lines. This workshop brought together over 100 scientists, from 13 countries, who are working in the academic and commercial research sectors, including experts and opinion leaders in the fields of embryology, animal imaging, data capture, quality control and annotation, high-throughput mouse production, phenotyping, and reporter gene analysis. This article summarises the outcome of the workshop, including (1) the vital scientific importance of phenotyping embryonic lethal mouse strains for basic and translational research; (2) a common framework to harmonise international efforts within this context; (3) the types of phenotyping that are likely to be most appropriate for systematic use, with a focus on 3D embryo imaging; (4) the importance of centralising data in a standardised form to facilitate data mining; and (5) the development of online tools to allow open access to and dissemination of the phenotyping data.
    Disease Models and Mechanisms 03/2013; 6(3). DOI:10.1242/dmm.011833 · 5.54 Impact Factor
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    • "Optical coherence tomography (OCT) allows for noninvasive, fast, single-cell resolution imaging through several millimeters of biological tissue. OCT, with its ability to provide accurate information on structure and dynamics, has been used for studying whole mammalian and avian embryos [1], [2], understanding hemodynamics [3], [4], and capturing 3D+time cardiac images in mammalian and avian embryos [5]–[7]. Frame rates currently achievable with direct, 3D+time OCT are too low to properly capture the dynamics of fast moving cardiovascular structures such as the beating embryonic heart. "
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    ABSTRACT: Current methods to build dynamic optical coherence tomography (OCT) volumes of the beating embryonic heart involve synchronization of 2D+time slice-sequences acquired over separate heartbeats. Temporal registration of these sequences is performed either through gating or post-processing. While synchronization algorithms that exclusively rely on image-intrinsic signals allow forgoing external gating hardware, they are prone to error accumulation, require operator-supervised correction, or lead to non-isotropic resolution. Here, we propose an imagebased, retrospective reconstruction technique that uses two sets of parallel 2D+T slice-sequences, acquired perpendicularly to each other, to yield accurate and automatic reconstructions with isotropic resolution. The method utilizes the similarity of the data at the slice intersections to spatio-temporally register the two sets of slice sequences and fuse them into a high-resolution 4D volume. We characterize our method by using (1) simulated heart phantom datasets and (2) OCT datasets acquired from the beating heart of live cultured E9.5 mouse and E10.5 rat embryos. We demonstrate that while our method requires greater acquisition and reconstruction time compared to methods that use slices from a single direction, it produces more accurate and self-validating reconstructions since each set of reconstructed slices acts as a reference for the slices in the perpendicular set.
    12/2012; 32(3). DOI:10.1109/TMI.2012.2231692
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