OPT produces high-resolution images (down to 6 μm) that can be improved twofold by postprocessing33 and can even be used in vitro in living tissue.34, The use of OPT for investigating anatomical structures in mouse embryos has been continuously improved and has led to astonishing results, such as the visualization of the whole vascular system of a developing mouse embryo after whole mount immunostaining with a PECAM1 antibody.35 Whole mount immunohistochemistry to label certain subsets of cells can be done in embryonic to larval stages in zebrafish36 but to use these methods to study the mechanism of tooth replacement in 1-month-old zebrafish, the protocols need to be adjusted, taking, for example, those established for mouse embryos as a starting point.37 While OPT has been utilized to make an anatomical atlas of zebrafish development,38 OPT is only a sporadic tool in zebrafish research (Fig. Zebra jaw bown in Resin, Tell: 1-800-351-8900 Steyer et al.17 took the HREM method one step further and developed an automatic cryo-imaging system from which fluorescence data can be obtained. By linking the movement of the microtome with the capturing of the image, whole blocks can be automatically cut and imaged.14. A solution of phosphotungstic acid in ethanol after Bouin's fixation gives good contrast in muscles, nervous tissues, and collagen-rich tissues (Fig. 4D) and related techniques have opened up a new direction in in vivo imaging. Image of amber, dentition, beautiful - 192042627 Smaller magnifications can be combined with better cameras to obtain high-resolution images. tiger's teeth aka canine teeth can grow up to 4-5 inches long depending on its size. Replacement teeth that have not reached the cytodifferentiation stage are, thus, not detected. 3C).26 Gallocyanin, a histological stain for nuclei, also gives good micro-CT contrast.27 The latter stain could potentially be used to detect epithelial thickenings and mesenchymal condensations. Ahrens MB, Orger MB, Robson DN, Li JM, Keller PJ. Teeth constantly renew, forming from an epithelial outgrowth called the successional lamina that is attached to the epithelial crypt of the predecessor tooth. Everyday. Heritable and precise zebrafish genome editing using a CRISPR-Cas system, Second harmonic generating (SHG) nanoprobes for. TPEF microscopy, however, has its drawbacks. The mountain zebra, an endangered species, also has a maximum of 40 teeth. If your smile is not becoming to you, then you should be coming to me! Henning AL, Jiang MX, Yalcin HC, Butcher JT. Photo about An adult tiger with open mouth growling and clear view of large fang teeth. Effect of excitation wavelength on penetration depth in nonlinear optical microscopy of turbid media, Two-photon imaging to a depth of 1000 in living brains by use of a Ti: Al2o3 regenerative amplifier, Highly nonlinear photodamage in two-photon fluorescence microscopy. Cryo-imaging of fluorescently labeled single cells in a mouse. One example is to use an antibody against basal lamina proteins such as laminin, or the alpha3(IV) chain of type IV collagen. Second, Bessel beams posses the ability to “self-heal,” which means that they can be partially obstructed at one point, but will reform after that point.61 The properties of Bessel beam illumination and widefield 3D super-resolution structured illumination microscopy62 have been used to image with a resolution under the diffraction limit in thickly fluorescent specimens.63 This Bessel-plane SR-SIM setup enabled high-speed, high 3D-resolution imaging with low-phototoxicity and an improvement in resolution of about 1.5× in the lateral resolution and 2× axially compared with confocal microscopy.64 Combining the Bessel-plane SR-SIM with two photon excitation resulted in high 3D-resolution images of the brain of a Drosophila melanogaster fly and a fixed Caenorhabditis elegans L1-stage larva. It uses these teeth in rivalry fights with other stallions. At the appointed time, Zebra arrived at Baboon’s home. Intensifying the exciting light will only result in an intensification of the emitted SHG waves. Studies on animals that replace their teeth repeatedly have revealed significant differences between the development of first-generation and replacement teeth, at least in teleosts and amphibians.1,2 Gaining a full understanding of continuous tooth replacement thus demands the use of a non-mammalian, easily accessible experimental model. Zebra skirmishes are frequent, and a serious zebra fight can be deadly. While light sheet microscopy has so far not been implemented in dental research, the various techniques under this banner are included here because they provide the current apex for deep and in vivo imaging. Does the male or female zebra have pointed canine teeth?, True or False Grevy Zebras live in the plains of Africa?, Do Grevy Zebras stay in herds or stay by themselves?, Is there breeding September through April or May through August? Color images available online at www.liebertpub.com/zeb. For example, the fibroblast growth factor-inhibitor SU5402 ablates dlx2b expression in the pharyngeal epithelium.11 Investigating the precise effect of SU5402 on the process of tooth replacement using the dlx2b:GFP line, combined with vibratome sections and CLSM, can therefore not be done. Anderson GA, Wong MD, Yang J, Henkelman RM. Amphibian teeth: current knowledge, unanswered questions, and some directions for future research. Listed in Folders. . Zebras evolved among the Old World horses within the last 4 million years. Zebra jaw bone in Resin, Supernatural : Fossil Series. S2. No they do not. Canis is a genus of the Caninae containing multiple extant species, such as wolves, dogs, coyotes and jackals.Species of this genus are distinguished by their moderate to large size, their massive, well-developed skulls and dentition, long legs, and comparatively short ears and tails. The fight was brutal with neither animal backing down. The axis of the detection objective is placed orthogonal to this light sheet, and the illuminated plane coincides with the focal plane of the detection objective. Using this setup, the heart of a 5.4-day-old zebrafish was imaged at a cellular resolution and a speed of 70 frames/s.60, Another way to enable better resolution at greater imaging depth is the use of Bessel beam illumination. For example, iodine potassium iodide after formalin fixation gave good soft tissue contrast in fry of Esox lucius (Fig. 1A), but are time-consuming. High-resolution episcopic microscopy: a rapid technique for high detailed 3D analysis of gene activity in the context of tissue architecture and morphology. . Boot MJ, Westerberg CH, Sanz-Ezquerro J, Cotterell J, Schweitzer R, Torres M, et al. In TPEF microscopy, the incident energy is absorbed so that a real excited state can be reached from which the fluorescence signal can emerge. Furthermore, the use of longer wavelengths reduces but does not eliminate photoxicity and light scattering44 and makes deep-tissue, in vivo imaging possible. Analyzing bone, blood vessels, and biomaterials with microcomputed tomography, Identification of putative dental epithelial stem cells in a lizard with life-long tooth replacement, Molecular dissection of craniofacial development using zebrafish. It stains not only calcified cartilage and mineralized bone matrix but also mineralized dentine (Fig. 4A). Figure 4C presents a preliminary attempt to obtain an in vivo view of the ventral tooth row of a 29-day-old zebrafish at a depth of 500 μm using SHG microscopy. Finally, by using 2P-SPIM over 2P-LSM, photodamage is decreased, which enables an increase in excitation power and thus an increase in signal-to-noise ratio and acquisition speed. . The name molar derives from Latin, molaris dens, meaning “millstone tooth”, from “mola”, millstone and dens, tooth. However, molecular evidence supports zebras as a monophyletic lineage. The maxillary arcade or upper jaw is listed over the mandibular arcadeor lower jaw. The incisors are indicated first, followed by the canine, promolars and molars. 4B).39 On the other hand, the difference in fluorescence lifetime in OPT can be used to distinguish between autofluorescence (which is especially strong in the yolk sac of developing zebrafish) and the fluorescence of GFP in a lysC:GFP zebrafish line.40, Combining two imaging modalities can, of course, greatly increase the information obtainable from the images. punk wasteland zebra female edgy grin teeth canine hybrid commission comm com open boss ass bitch. The setup was used to record 80% of the neurons in the brain of a 5-day-old GCaMP5G zebrafish line at single cell resolution.59 The entire larval zebrafish brain, covering a volume of 800×600×200 μm, was imaged in approximately 1.3 s. In addition, light sheet microscopy has been combined with two-photon fluorescence (2P-SPIM) to extend the imaging depth of one-photon light sheet microscopy by a factor of two.58,60 Furthermore, the axial resolution of 2P-SPIM achieved at higher imaging depth is superior to both one-photon light sheet and two photon laser scanning microscopy (2P-LSM) but at greater imaging depth the lateral resolution of 2P-LSM is better than that of 2P-SPIM. All these teeth end evenly, so that the zebra can easily clip grass. Fish were sedated with 0.016% MS-222 and recovered completely after 20 min of measurements. Gill arch and hyoid arch diversity and cypriniform phylogeny: distributed integration of morphology and web-based tools, Dynamics of tooth formation and replacement in the zebrafish (, Formation of a successional dental lamina in the zebrafish (. (E) Three-dimensional (3D) reconstruction of the sectioned zebrafish with the dentition and a part of the pharyngeal arches highlighted, right view shows details of the 3D-reconstruction, scale bar left: 200 μm. Whole-brain functional imaging at cellular resolution using light-sheet microscopy. Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain. M.M. S1, Supplementary Methods and Supplementary Fig. 61 mil gostos. Shown is a view of kidney cellular structure with podocytes labeled with mCherry and endothelial cells labeled with green fluorescent protein (GFP). To ameliorate in vivo SHG microscopy, BaTiO3 nanocrystals can be used as a contrast enhancer.53 These BaTiO3 nanoprobes do not suffer from the aforementioned problems of fluorescent molecules. For example, using the properties of 10% potassium triiodide or Lugol solution and optimizing the staining technique to avoid tissue shrinkage, the developing cardiovascular system could be tracked in mouse embryos from E10.5 to E17.5.25 The use of lugol and other soft tissue contrast enhancing agents has also been described in great detail for other species. So far, many of these possibilities either have been explored only in the most basic way or have not even been touched upon. The combination of such a pulsed laser with an objective with a high numerical aperture forces a large number of photons to the region of interest. Since the probability that two photons are interacting with the same molecule at the same time is very small, high-power, pulsed lasers must be used as the source of excitation. Tissue architecture can be distinguished by the varying ability of the different cell types to hinder the fluorescence of the embedding medium (Fig. . 25 baby zebra clipart on PN cliparts. Zu Y, Tong X, Wang Z, Liu D, Pan R, Li Z, et al. S1 (Supplementary Data are available online at www.liebertpub.com/zeb). Email: contact@mercorne.fr Email: jantz@jantzusa.com 2F). In the Alizarin complexone form, Alizarin red can also be used for in vivo staining (Fig. Just one more reason to smile! Their kicks are tremendously powerful, and the males have erupted canine teeth that they use to bite their opponents – broken skin and bones are not uncommon, and many a zebra have lost their tail as a result of a fight. We finally discuss methods based on technological developments that allow us to visualize tooth replacement in vivo, and, to some extent, enable observations on soft tissues as well. It’s all about you. HREM-method is outlined in the Supplementary Methods and Supplementary Fig. While many also possess a few molars in the back of their mouths, and sharp incisors in the front, the most important teeth for carnivores are their long, sharp canine teeth. Helping keep Portland’s kids smiling! Guldberg RE, Ballock RT, Boyan BD, Duvall CL, Lin ASP, Nagaraja S, et al. SHG method is outlined in the Supplementary Methods and Supplementary Fig. . Wu P, Wu XS, Jiang TX, Elsey RM, Temple BL, Divers SJ, et al. The zebrafish, a model organism for which a plethora of molecular and genetic techniques exists, has a lifelong replacing dentition of 22 pharyngeal teeth. Most carnivores have long, sharp teeth adapted to ripping, tearing or cutting flesh. zebrafish (courtesy of D. Adriaens), scale bar: 1 mm. Zhao M, Zhang H, Li Y, Ashok A, Liang R, Zhou W, et al. It’s time to smile again. Zebra Molars in Resin, Supernatural : Fossil Series. This SIMview setup obliterates the need for time-consuming sample rotation. 1E, F). (B) Three-dimensional-reconstruction of flow-OPT data, showing the blood vessels in the trunk of a juvenile casper zebrafish, reprinted by permission from Bassi et al.39 First-generation teeth in nonmammalian lineages: evidence for a conserved ancestral character? Physical Address Mercorne, ZI, 48300 Langogne, France, Tell: (02) 9648 5856 This eliminates the need for a confocal pinhole configuration, and non-descanned detection can be used. Teeth constantly renew, forming from an epithelial outgrowth called the successional lamina that is attached to the epithelial crypt of the predecessor tooth.7 The successional lamina thickens in the early morphogenesis stage, re-invaginates, and forms a bi-layered bell-shaped structure, consisting of an outer and an inner epithelial layer, during the late morphogenesis stage. For instance, a dog has 3 incisors, 1 canine, 4 premolars and 2 molars on one side of the upper jaw and 3 incisors, 1 canine, 4 premolars and 3 molars on one side of the lower jaw, so the de… (A) Micro-computed tomography scan of head skeleton and dentition of a 3.6 cm T.L. Zebra Molars and jaw bone in Resin, Supernatural : Fossil Series. Techniques that can circumvent this time- and labor-consuming routine by visualizing the dentition in vivo or at least in toto thus offer great advantages. would like to thank Lukas Reissig for his helpful comments. All zebras are dark-skinned animals. Because the jaw is bilaterally symetrical, only one half of the jaw is described. Bassi A, Fieramonti L, D'andrea C, Mione M, Valentini G. Mcginty J, Taylor HB, Chen L, Bugeon L, Lamb JR, Dallman MJ, et al. These slices can then be optically sectioned using confocal laser scanning microscopy (CLSM). Bart Bruneel, Markus Mathä, [...], and Ann Huysseune. FishFace: interactive atlas of zebrafish craniofacial development at cellular resolution, Multilayer mounting for long-term light sheet microscopy of zebrafish, Supplementary Methods and Supplementary Fig. Krzic U, Gunther S, Saunders TE, Streichan SJ, Hufnagel L. Multiview Light-sheet microscope for rapid, Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy. . In a synchrotron light source, electrons are accelerated close to the speed of light and kept inside the circular setup. A DAPI stain for nuclei, a phalloidin stain for F-actine,4 or a general vital stain, such as bodipy ceramide, can be utilized to provide the necessary counterstain (Fig. Noninvasive imaging beyond the diffraction limit of 3D dynamics in thickly fluorescent specimens. Finally, the possibilities of light sheet microscopy are addressed. Multiphoton autofluorescence and second-harmonic generation imaging of the tooth. Degenhardt K, Wright AC, Horng D, Padmanabhan A, Epstein JA. Between the fixation of 1-month-old zebrafish and the eventual visualization with conventional light microscopy, it takes on average 2–3 weeks for decalcification, postfixation, embedding, sectioning, staining, and mounting. ), caudal view, scale bar: 0.3 mm. A coming of age: advanced imaging technologies for characterizing the developing mouse. For example, the use of a hybrid fluorescence molecular tomography/micro-CT system to combine micro-CT data on anatomical structures with functional fluorescent reporter gene expression patterns in mice has been reported.41. (B) Transverse virtual section of formalin-fixed and iodine potassium iodide-stained Esox lucius fry, reprinted by permission from Metscher.27 Helping you smile. Walls JR, Sled JG, Sharpe J, Henkelman RM. In this way, HREM imposes no constraints on the size of the tissue sample.16 All these advantages make HREM a fast and easy method to visualize samples on a cellular level. First, a perfect Bessel beam does not diffract, which means a light sheet formed by a Bessel beam will only spread out minimally and retain its initial thickness. The Teeth of Herbivores, Carnivores and Omnivores. Quantitative three-dimensional imaging of live avian embryonic morphogenesis via micro-computed tomography. Furthermore, they have a narrow scatter spectrum, making them distinguishable from autofluorescence, and they have an SHG signal that is 10 times stronger than the signal of endogenous material such as muscle fibers. Deep and fast live imaging with two-photon scanned light-sheet microscopy, Optical sectioning microscopy with planar or structured illumination. A global study of ancient dog DNA presents evidence that there were different types of dogs more than 11,000 years ago in the period immediately following the Ice Age. Rapid 3D Phenotyping of cardiovascular development in mouse embryos by micro-CT with iodine staining, MicroCT for comparative morphology: simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues, MicroCT for Developmental biology: a versatile tool for high-contrast 3D imaging at histological resolutions. (D) Light sheet microscopic image showing the vasculature in a 78-hour postfertilization (hpf) Tg(kdrl:GFP) transgenic zebrafish, scale bar: 125 μm, reprinted by permission from Weber et al.68 Once the stage of late cytodifferentiation is reached, the deposition of the enameloid and dentin matrix at the tooth tip begins. Male Zebras have pointed canine teeth usually only found on meat eaters. 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zebra canine teeth

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