Live optical projection tomography

Centre for Genomic Regulation
Organogenesis (Impact Factor: 2.8). 10/2009; 5(4):211-6. DOI: 10.4161/org.5.4.10426
Source: PubMed


Optical projection tomography (OPT) is a technology ideally suited for imaging embryonic organs. We emphasize here recent successes in translating this potential into the field of live imaging. Live OPT (also known as 4D OPT, or time-lapse OPT) is already in position to accumulate good quantitative data on the developmental dynamics of organogenesis, a prerequisite for building realistic computer models and tackling new biological problems. Yet, live OPT is being further developed by merging state-of-the-art mouse embryo culture with the OPT system. We discuss the technological challenges that this entails and the prospects for expansion of this molecular imaging technique into a wider range of applications.

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Available from: jean-francois Colas, May 10, 2014
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    • "As a consequence, the overall resolution and contrast in the OPT reconstructions are compromised. Moreover, SPIM is considered an ideal technique to image development in vivo, whereas tomographic imaging is commonly believed suitable only for fixed and chemically cleared samples and, indeed, only a few in vivo applications have been reported (Colas and Sharpe, 2009; Bassi et al., 2011; McGinty et al., 2011). We decided to look for alternative solutions that would achieve tomographic reconstructions of living specimens in a highresolution , high-NA SPIM setup without any modifications to the hardware. "
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    ABSTRACT: Fluorescently labeled structures can be spectrally isolated and imaged at high resolution in living embryos by light sheet microscopy. Multimodal imaging techniques are now needed to put these distinct structures back into the context of the surrounding tissue. We found that the bright-field contrast of unstained specimens in a selective plane illumination microscopy (SPIM) setup can be exploited for in vivo tomographic reconstructions of the three-dimensional anatomy of zebrafish, without causing phototoxicity. We report multimodal imaging of entire zebrafish embryos over several hours of development, as well as segmentation, tracking and automatic registration of individual organs. © 2015. Published by The Company of Biologists Ltd.
    Development 02/2015; 142(5). DOI:10.1242/dev.116970 · 6.46 Impact Factor
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    • "The second properties of infrared light, the infrared light can't be seen by naked eyes but it can be seen by using the digital camera. The optical tomography system is suitable for use on bio tissue and non-bio tissue [9] [14] [15] [16] [17]. However, the size of the subject must be considered first, as the subject must not be too small or too large. "
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    ABSTRACT: In this paper we use an electronic component to produce light which is applied in testing soft tissue penetration. We used bio tissue, a slice of apple, and non-bio tissue, paper. The voltage could be adjusted to brighten the light to view the penetration of the subject. The thickness of the tissue was constant and the results showed that the current and voltage were significant as the light penetrated the soft tissue.
    09/2014; 70(3). DOI:10.11113/jt.v70.3475
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    • "The combination of visualizing gene expression throughout the whole volume of mouse embryos and overall morphology with OPT proves to be essential for studies in neural [37], [38] and lung [39]–[42] development as well. OPT also has the potential to investigate in-vivo processes [43]. Modifications in hardware and the development of sophisticated software algorithms [44], [45] have made real-time visualization of internal structures possible, such as the skeletal and nervous systems of the zebrafish [46]. "
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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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