Dr. Jeremy Logue. The ventricular or apical surface is adjacent to the retinal pigment epithelium (pe), and the basal surface is adjacent to the lens (le). x����Z$�v��,���i�� �j��Yp�nh�� ���Rȳ�f?��P�F ���+X�!��u��N����*P������!+32"2��U ��c�^/�M����p���'߻7�Q��C�勑l�����:��O6��|I2]n�j直�=. Zebrafish have long been utilized to study the cellular and molecular mechanisms of development by time-lapse imaging of the living transparent embryo. Zebrafish Core in the NICHD Annual Report Annotated zebrafish development timelapse. Time-lapse image recording reveals that nephrogenesis in control morpholino injected embryos is unaltered in comparison to uninjected ones (results not shown) and follows the described steps of early zebrafish kidney development 3. In zebrafish, the lens placode appeared in the head ectoderm, similar to mammals. Zebrafish Embryogenesis Brightfield imaging Time-lapse Morphogenetic movements Population statistics Quantification Temperature control This is a … Time-lapse movie of zebrafish lens epithelial cells expressing GFP-histones and mCherry-zGem as they progress through the cell cycle. Summary. Two-photon excitation microscopy was used to reconstruct cell divisions in living zebrafish embryonic retinas. 5 0 obj Edinburgh, Scotland . Time-lapse imaging was performed using a Leica TCS SP8 upright microscope with a Leica HCX IRAPO L ×25/0.95 water-dipping objective and heating chamber, or on an upright 3i spinning-disc confocal using a Zeiss Plan-Apochromat 20×, 40× or 63×/1.0 NA water-dipping objective. stream 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. Still image of the spinal cord from time lapse imaging of a 46 hours post-fertilization embryo harboring a transgenic reporter marking sox10+ cells. Confocal. This is particularly useful for following cellular interactions among like cells, which are difficult to label differentially using traditional promoter-driven colors. 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. /Im2 9 0 R /Im3 12 0 R >> >> Annotated zebrafish development timelapse. xڭ�]o�0F��+���"�? The Roslin Institute. 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- 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. Although traditionally used for developmental biology, zebrafish has recently been used to investigate metabolic diseases. 2 Imaging Station - Formative Evaluation 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- An efficient method for isolating embryonic zebrafish neurons from neural tissue. The video was captured for the first time by a new microscope imaging technique part-funded by the BHF. %��������� This specimen, stained with Bodipy-FL-C 5-Ceramide, shows the outline of all cells in the retinal neuroepithelium (ne). interval over a period of 13 h (Additional file 1: Movie S1). Schematic diagram of the apparatus used for long-term time-lapse imaging of developing zebrafish. Time-Lapse Analysis of Cell Division in the Zebrafish Embryonic Retina. However, our time-lapse imaging indicates that, in zebrafish, microglial precursors appear to enter the optic tectum directly from the periphery . Using in vivo lineage tracing by time-lapse confocal microscopy combined with morpholino oligonucleotide-mediated gene inactivation, we report the function of FGF3 during zebrafish POMC cell ontogeny, defining a genetic and developmental boundary between two distinct pituitary POMC lineages in a gene-dose-dependent fashion. Early in the time-lapse video, cells can be seen dividing with little change in their shape or appearance. Annotated zebrafish development timelapse is shown in this video. Fluorescence. Compound transgenic fish … Swinburne IA, Mosaliganti KR, Green AA, Megason SG. << /Length 6 0 R /Type /XObject /Subtype /Image /Width 1800 /Height Indeed, time-lapse imaging showed that EHT occur in the ventral wall of CA from about 32 hpf until 72 hpf (Fig. You need to login to download this video. This process have these following stepys. 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. 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 In vivo, high‐resolution, time‐lapse imaging characterized lens development in the zebrafish from 16 to 96 hr postfertilization (hpf). The upper part shows… Here, we report the … 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。 Researchers from the universities of Glasgow and Edinburgh were able to record individual heart cells growing and dividing in … Or press "Snap Photo" again to take another image! Enterprise . Time lapse imaging was conducted for a period of 14–17 h with the time interval between images set at 3 min. 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. This process have these following stepys. Time-lapse images of a zebrafish spinal neuron growth cone in culture loaded with a fluorescent calcium sensor. Adaptive prospective optical gating enables day-long 3D time-lapse imaging of the beating embryonic zebrafish heart, … At time = 0∶00 min, BDNF is applied in the bath and initiates a Ca 2+ signal in the growth cone by 5 min. More information: Jonathan M. Taylor et al. In vivo time-lapse imaging of cell divisions during neurogenesis in the developing zebrafish retina. 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. Extended time-lapse imaging of vascular, neuronal and muscle development After optimizing the mounting method described above, time lapse confocal microscopy images were captured over a span of 55 h. We imaged live transgenic zebrafish expressing GFP or RFP in different tissues. ( Send Message ) on 10-12-2007. *To avoid redundant efforts, please check whether a suitable zebrafish line is already available through existing databases. endstream Dr. Nils Lindstrom. endobj Somitogenesis in the zebrafish embryo is shown in Movie 1 . Time-lapse imaging of zebrafish embryos is also described by Köster and Fraser (2004) and Distel et al. We analysed time-lapse movies of chevron formation during zebrafish development at 28°C, which were acquired in our previous studies (Herrgen et al., … (2006). Uploaded by: Assessment of heart function via time‐lapse video microscopy in Zebrafish. • Primary cells can survive 9 days in vitro, expressing markers for neurons and glia. The acquired z stacks for both single time point and time lapse images were selected to encompass all regions of interest within the zebrafish larval brain. << /ProcSet [ /PDF /Text /ImageB /ImageC /ImageI ] /ColorSpace << /Cs1 Unit or Division: Developmental Neurobiology Unit (Ichiro Masai) Free for anyone to re-use, but must be credited to OIST. 3, K–P; and Video 1). login or signup, Channels: Boxes designate the ventricle and dorsal aorta regions. Improved long-term imaging of embryos with genetically encoded α … Tubing carrying water is … Because of the rapid embryogenesis, external development, and transparency of zebrafish embryos, their developmental processes can be visualized in time-lapse studies in the context of the living organism. 1 0 obj Here, we report the use of confocal Raman spectroscopic imaging for … A mounting method for extended time-lapse confocal microscopy of whole zebrafish embryos is described here. Field of Research: Developmental Neuroscience. Time-lapse movie of zebrafish lens epithelial cells expressing GFP-histones and mCherry-zGem as they progress through the cell cycle. stream Zebrafish larvae were treated at 48 hpf with SKP2-C25 (1% DMSO). In zebrafish, the lens placode appeared in the head ectoderm, similar to mammals. Neuron 37 , 597 - 609 . [0 0 612 792] >> 2009;546:243–54. 100 /ColorSpace 7 0 R /BitsPerComponent 8 /Filter /FlateDecode >> Right click on the image below and then select "Save image as" option to save the image for your powerpoint presentations. Materials and methods A: Brightfield image of a 3‐dpf transparent Zebrafish embryo. Developing mouse embryonic kidneys . * Interoperability. 3 0 obj Zebrafish possess many advantages that make them the best vertebrate model organism for live imaging of dynamic development events. %PDF-1.3 4x. By this time, the basic vertebrate body plan is recognizable and several organ systems are … This process have these following stepys. adult formation. 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. Process remodeling and migration paths are highly variable and seem to be influenced by contact with neighboring OPCs. 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. endobj National Institutes of Health. Methods Mol Biol. 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 time-lapse imaging Tg(sox10:mRFP; olig2:GFP) larvae were used. 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. 4 0 obj << /Length 4 0 R /Filter /FlateDecode >> The timelapse movie shows a developing zebrafish embryo Tg(kdrl:GFP) from 24-96 hpf imaged using selective plane illumination microscopy (SPIM). 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. This article describes a layered mounting method for zebrafish embryos that restrict the motility of the embryos while allowing for the unrestricted growth. 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. Others. Time-lapse FRET imaging. In this study, using the zebrafish as the model system, we attempted to answer this classic question. Precise patterns of division, migration and differentiation of neural progenitor cells are crucial for proper brain development and function 1,2.To understand the behavior of neural progenitor cells in the complex in vivo environment, time-lapse live imaging of neural progenitor cells in an intact brain is critically required. 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. Multiple embryo time-lapse imaging of zebrafish development. Contrary to proposed models for vertebrate asymmetric divisions, no apico-basal cell divisions take place in the zebrafish retina during the generation of postmitotic neurons. gastrula stage, 24 hour, one day, two day, three day embryo, newly hatched three-day fish and 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 … Cryopreservation of transgenic and mutant zebrafish lines; Importation of existing zebrafish lines; Characterization of phenotypes by time-lapse photography Get started. scitan We demonstrate the utility of vascular-specific transgenic zebrafish in conjunction with time-lapse multiphoton laser scanning microscopy by directly observing angiogenesis within … The optical transparency of its embryos permits time lapse live imaging (Hall et al., 2009; Herrgen et al., 2009; Feierstein et al., 2015). doi: 10.1016/S0896-6273(03)00066-7 OpenUrl CrossRef PubMed Web of Science 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. 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