2 0 obj Time-lapse images of a zebrafish spinal neuron growth cone in culture loaded with fluorescent Ca 2+ sensor Fluo-8H (pseudocolored according to scale in Figure 6A). Rapid embryonic development A major benefit of using the zebrafish as a model is its rapid early development. Figure 2c shows three timepoints of a single time-lapse recording with 34 positions, 2 stitched FOVs, 2 channels (transgenes cldnb:GFP and cxcr4b:H2B-RFP), imaged with a 10 min. 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. For Core Members Zebrafish possess many advantages that make them the best vertebrate model organism for live imaging of dynamic development events. Heterogeneity of stem cells or their niches is likely to influence tissue regeneration. Intravital time-lapse imaging is a powerful technique for investigating continuous developmental processes without missing crucial events. 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. This process have these following stepys. xڭ�]o�0F��+���"�? While mounting other fish check back to make sure previous subjects maintain desired orientation until agarose solidifies (~3 min). Boxes designate the ventricle and dorsal aorta regions. We describe a new imaging chamber that provides continuously circulating flow of warm, oxygenated aqueous media. 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. doi: 10.1016/S0896-6273(03)00066-7 OpenUrl CrossRef PubMed Web of Science In vivo time-lapse imaging of cell divisions during neurogenesis in the developing zebrafish retina. Time-lapse movie of zebrafish lens epithelial cells expressing GFP-histones and mCherry-zGem as they progress through the cell cycle. 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. [0 0 612 792] >> The video was captured for the first time by a new microscope imaging technique part-funded by the BHF. 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. Developing mouse embryonic kidneys . ( Send Message ) on 10-12-2007. Time-lapse imaging of zebrafish embryos is also described by Köster and Fraser (2004) and Distel et al. The optical transparency of its embryos permits time lapse live imaging (Hall et al., 2009; Herrgen et al., 2009; Feierstein et al., 2015). The mounting is performed in layers of agarose at different concentrations. Early in the time-lapse video, cells can be seen dividing with little change in their shape or appearance. *To avoid redundant efforts, please check whether a suitable zebrafish line is already available through existing databases. 4x. Process remodeling and migration paths are highly variable and seem to be influenced by contact with neighboring OPCs. 3 0 obj We demonstrate the utility of vascular-specific transgenic zebrafish in conjunction with time-lapse multiphoton laser scanning microscopy by directly observing angiogenesis within … Assessment of heart function via time‐lapse video microscopy in Zebrafish. interval over a period of 13 h (Additional file 1: Movie S1). Time-lapse of a 2 day old zebrafish embryo with green erythrocytes and red blood vessels. The Aquatic Zebrafish Core provides services using the small vertebrate zebrafish as a model for human disease and to study gene function. Here we reveal stem/precursor cell diversity during wound repair in larval zebrafish somitic body muscle using time-lapse 3D confocal microscopy on reporter lines. 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 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). Delamination of the surface ectoderm resulted in the formation of the lens mass, which progressed to a solid sphere of cells separating from the developing cornea at approximately 24 hpf. To determine whether the Erk biosensor could be used to monitor Erk activity in zebrafish embryos, 8SS embryos carrying the Erk biosensor were observed for 60 or 75 min at 5 min intervals at 28.5 °C. B: M‐mode imaging through short axis of the ventricle. Zebrafish Core in the NICHD Annual Report Zebrafish larvae were treated at 48 hpf with SKP2-C25 (1% DMSO). 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