Boxes designate the ventricle and dorsal aorta regions. We analysed time-lapse movies of chevron formation during zebrafish development at 28°C, which were acquired in our previous studies (Herrgen et al., … Swinburne IA, Mosaliganti KR, Green AA, Megason SG. Time-Lapse Video of Zebrafish “Inner Ear” Development Wins Small World in Motion Competition Posted On May 5, 2015 Nikon Instruments Inc. recently announced the winners of the fourth annual Nikon Small World in Motion Photomicrography Competition. For example, the time lapse movie to the right depicts the first 18 hours of development. The transparency and rapid embryogenesis of zebrafish, together with fluorescent reporters, allow for the study of developmental processes with single-cell resolution using in vivo time-lapse imaging (Beis and Stainier, 2006). This largely results from the fact that zebrafish embryos are transparent and thus accessible to various imaging techniques, such as confocal and two-photon excitation (2PE) microscopy. Time-lapse movie of zebrafish lens epithelial cells expressing GFP-histones and mCherry-zGem as they progress through the cell cycle. Research methodologies include the following: gene expression studies, gain or loss of gene function studies, cryopreservation of zebrafish lines, importation of new lines, and characterization of phenotypes by time-lapse photography. Zebrafish Core in the NICHD Annual Report Zebrafish larvae were treated at 48 hpf with SKP2-C25 (1% DMSO). A: Brightfield image of a 3‐dpf transparent Zebrafish embryo. 1 cell, 2 cell, 4 cell, 8 cell, early blastula, mid-blastula, late blastula, gastrula stage, 24 hour, one day, two day, three day embryo, newly hatched three-day fish and adult formation. A time-lapse 3D video of a zebrafish heart growing over a day has been captured for the first time by a new microscope imaging technique. This article describes a layered mounting method for zebrafish embryos that restrict the motility of the embryos while allowing for the unrestricted growth. * Interoperability. Skeletal muscle with incision wounds rapidly regenerates both slow and fast muscle fibre types. A mounting method for extended time-lapse confocal microscopy of whole zebrafish embryos is described here. This is particularly useful for following cellular interactions among like cells, which are difficult to label differentially using traditional promoter-driven colors. Rapid embryonic development A major benefit of using the zebrafish as a model is its rapid early development. endobj For serial time-lapse experiments (see below) we typically mount six fish at a time and try to limit imaging sessions to 1 hour per group. Zebrafish possess many advantages that make them the best vertebrate model organism for live imaging of dynamic development events. Time-lapse movie of zebrafish lens epithelial cells expressing GFP-histones and mCherry-zGem as they progress through the cell cycle. There are numerous reasons to model a disease in a fish, including the rapid, five-day development of zebrafish and the ability to image whole organs in vivo and perform time-lapse analysis. One of the most important questions is whether the positional information for the pattern formation is derived from a covert prepattern or an autonomous mechanism. Here we describe a technique for using time-lapse confocal microscopy to visualize large numbers of multicolor Brainbow-labeled cells over real time within the developing zebrafish nervous system. Fluorescence. The timelapse movie shows a developing zebrafish embryo Tg(kdrl:GFP) from 24-96 hpf imaged using selective plane illumination microscopy (SPIM). In this study, using the zebrafish as the model system, we attempted to answer this classic question. 4x. At time = 0∶00 min, BDNF is applied in the bath and initiates a Ca 2+ signal in the growth cone by 5 min. By this time, the basic vertebrate body plan is recognizable and several organ systems are … The optical transparency of its embryos permits time lapse live imaging (Hall et al., 2009; Herrgen et al., 2009; Feierstein et al., 2015). endobj 7 0 R >> /Font << /F1.0 8 0 R /F2.0 11 0 R >> /XObject << /Im1 5 0 R Annotated zebrafish development timelapse is shown in this video. Tubing carrying water is … For time-lapse imaging Tg(sox10:mRFP; olig2:GFP) larvae were used. Dr. Nils Lindstrom. ( Send Message ) on 10-12-2007. In zebrafish, our high-resolution time-lapse imaging allowed us to definitively determine that endoderm-derived cells of Seessel’s pouch detach … A variety of different transgenic zebrafish lines have been generated expressing green fluorescent protein (GFP) or other fluorescent proteins in different organs and tissues, permitting dynamic visualization of development of these organs and tissues in living animals via time-lapse … Enterprise . Uploaded by: Check out this time-lapse 3D video of a zebrafish heart growing over a day! In vivo time-lapse imaging of cell divisions during neurogenesis in the developing zebrafish retina. Apparatus for long-term time-lapse imaging. Annotated zebrafish development timelapse. In Vivo Time-Lapse Imaging of Zebrafish Embryonic Development -- Distel and Köster 2007 (16): pdb.prot4816 -- Cold Spring Harbor Protocols Reagent Amount to add 2.9 M NaCl 60 mL For Core Members We describe a new imaging chamber that provides continuously circulating flow of warm, oxygenated aqueous media. Here, we report the use of confocal Raman spectroscopic imaging for … Confocal. Methods Mol Biol. Time-Lapse Analysis of Cell Division in the Zebrafish Embryonic Retina. 4 0 obj Although methods have been devised for shortto medium-term time-lapse imaging of transgenic zebrafish, these methods are not suitable for longer term imaging because of poor control over temperature, evaporation, and anoxia. Early in the time-lapse video, cells can be seen dividing with little change in their shape or appearance. Intravital time-lapse imaging is a powerful technique for investigating continuous developmental processes without missing crucial events. • Here we describe a method to mount zebrafish embryos for long-term imaging and demonstrate how to automate the capture of time-lapse images using a confocal microscope. Zebrafish embryos provide a unique opportunity to visualize complex biological processes, yet conventional imaging modalities are unable to access intricate biomolecular information without compromising the integrity of the embryos. Denne video handler om Timelapse zebrafish. Dr. Jeremy Logue. This protocol describes a method to visualize clones of progenitor cells and neurons in the developing zebrafish hindbrain and follow them in vivo using Brainbow and time-lapse confocal microscopy 11.The major advantage of this protocol in comparison to in vitro or ex vivo studies is the ability to directly observe the proliferative zone of the vertebrate brain in its natural milieu over time. scitan In zebrafish, the lens placode appeared in the head ectoderm, similar to mammals. The most critical step for the mounting method is to identify the optimal concentration of agarose that will allow for unrestricted zebrafish embryo growth, and at the same time keep the embryos in a completely fixed position for confocal imaging. The upper part shows… doi: 10.1016/S0896-6273(03)00066-7 OpenUrl CrossRef PubMed Web … 1 cell, 2 cell, 4 cell, 8 cell, early blastula, mid-blastula, late blastula, gastrula stage, 24 hour, one day, two day, three day embryo, newly hatched three-day fish and adult formation. Summary. You need to login to download this video. ゼブラフィッシュの上皮細胞が、細胞周期の進行に伴いGFP-histoneとmCherry-zGemを発現する様子を示したタイムラプス動画。 To support this observation, we carried out time-lapse imaging analysis with Tg(kdrl:eGFP;coro1a:DsRedx) embryos in which the brain vessels and microglia are marked by GFP and DsRedx, respectively. endobj The mounting is performed in layers of agarose at different concentrations. The zebrafish system affords several significant advantages to these studies, including high-resolution in vivo time-lapse imaging and large-scale genetic and chemical screening. 2009;546:243–54. << /Type /Page /Parent 14 0 R /Resources 3 0 R /Contents 2 0 R /MediaBox Zebrafish Development Time-Lapse Video - Holding Time Joyce Ma August 2002 PURPOSE To determine how long visitors stay to watch the time-lapse video of Zebrafish development To determine if visitors stay to watch the entire video To determine if there is any difference in holding time between the annotated and the non- Edinburgh, Scotland . Time-lapse of a 2 day old zebrafish embryo with green erythrocytes and red blood vessels. While mounting other fish check back to make sure previous subjects maintain desired orientation until agarose solidifies (~3 min). Here we describe a technique for using time-lapse confocal microscopy to visualize large numbers of multicolor Brainbow-labeled cells over real time within the developing zebrafish nervous system. 2 0 obj At 56–57 hpf, larvae were anaesthetised in 600 μM tricaine and manually mounted in 1.5% low melting point agarose in embryo medium containing either SKP2-C25 (1% DMSO) or 1% DMSO and 600 μM tricaine. Using in vivo time-lapse confocal microscopy, we show that zebrafish OPCs continuously extend and retract numerous filopodium-like processes as they migrate and settle into their final positions. Adaptive prospective optical gating enables day-long 3D time-lapse imaging of the beating embryonic zebrafish heart, … The Aquatic Zebrafish Core provides services using the small vertebrate zebrafish as a model for human disease and to study gene function. An efficient method for isolating embryonic zebrafish neurons from neural tissue. 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