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400 mg tricaine powder 97.9 ml DD water ~2.1 ml 1 M Tris (pH 9). Adjust pH to ~7. Store this solution in the freezer. (Buy the smallest amount possible because tricaine gets old.) To use tricaine as an anesthetic combine the following in a 250 ml beaker: 4.2 ml tricaine solution ~100 ml clean tank water. TTBS: 20 mM Tris, pH 7.5 500 mM NaCl 0. Chapter 2 - Breeding Embryo Production By In Vitro Fertilization (Source: C. Walker and G. Streisinger) Overview of in vitro fertilization: Large numbers of synchronously developing embryos can be obtained by in vitro fertilization. Maintain breeding males and females on a daily schedule as described in the Zebrafish Breeding Schedule for Maximal Embryo Production (page 2.4) Without the tricaine, the fish are very sensitive to almost any stimulus and react with extremely strong startle responses. Without the tricaine step, a high percentage (50% or more) of mutagenized fish die. With the tricaine step, the death rate drops to almost zero. Prepare: ENU preparation buffer (fish system water, 10 mM PO4, pH 6.5) ZFIN is hiring both a software developer and a curator! The Zebrafish Information Network (ZFIN) is the database of genetic and genomic data for the zebrafish (Danio rerio) as a model organism.ZFIN provides a wide array of expertly curated, organized and cross-referenced zebrafish research data THE ZEBRAFISH BOOK. A guide for the laboratory use of zebrafish. Danio (Brachydanio) rerio. by Monte Westerfield, Institute of Neuroscience, University of Oregon. GENERAL METHODS FOR ZEBRAFISH CARE. BREEDING. EMBRYONIC AND LARVAL CULTURE. MICROSCOPIC OBSERVATIONS. CELLULAR METHODS

Tricaine methanesulfonate, also known as MS-222, is a synthetic local anesthetic commonly used by the fishing industry (Sato et al., 2000). It is the only anesthetic approved by the FDA (Food and Drug Administration) for use in fish intended for human consumption (Ross and Ross, 2008) Make the hole in the chorion bigger, little by little until it is big enough to release the embryo. 2. Pipette the embryo into a nine-well dish/a depression slide well containing fish water. 3. Use a pipette to transfer MS-222 or Tricaine drop-wise into wells. Anesthesia usually effective in less than a minute RINGER'S SOLUTIONS. SALT STOCK. SCHÖNEFELD'S MEDIUM FOR GROWING PARAMECIA. SDS SAMPLE BUFFER. SODIUM BICARBONATE. SODIUM THIOSULFATE SOLUTION 1.0%. SPERM EXTENDER. SPERM FREEZING MEDIUM. SPERM THAWING MEDIUM

Suck an embryo up into a Pasteur pipette and position it near the opening in the pipette. 3. Plunge the pipette into the methyl cellulose and gently expel the embryo with as little medium as possible. The medium around the embryo will be quickly absorbed into the methyl cellulose. 4. Use a fine loop of nylon fishing line to orient the embryo. 5 From zfin: Tricaine: Tricaine (3-amino benzoic acid ethyl ester also called ethyl 3-aminobenzoate) comes in a powdered form from Sigma (Cat.# A-5040). It is also available as Finquel (Part No. C-FINQ-UE) from Argent Chemical Laboratories, Inc. Make tricaine solution for anesthetizing fish by combining the following in a glass bottle with a. MS-222 (Tricaine Methanesulfonate) Preparation • Stock solution: 4 - 10 mg/ml - Do not buffer - Typically frozen at - 20 C and will be good for 3 months - Must label and date • Working solution: 0.2 mg/ml - Buffer solution to pH 7-8 with sodium bicarbonate - Typically refrigerated at 4 C and will be good for 1 month after creatio • Anesthesia with tricaine methane sulfonate (MS222, 168 mg/l) followed by rapid freezing in liquid nitrogen. 2. For zebrafish 4-7 days post-fertilization (dpf) the following methods are acceptable for euthanasia: • Immobilization by submersion in ice water (5 parts ice/1 part water, 0-4º C) for at least 2

ZFIN: Zebrafish Book: Breeding Zebrafis

Home Common Techniques Classroom Experiments Virtual Experiments Tutorials Games Glossary Links Publishing Opportunities About This Site Contact Us ZFIN 2.Euthanatize fish with an overdose of MS-222 (tricaine methane sulfonate). 3.Carefully slit open the body cavity along the belly, trying not to disrupt internal organs. 4.Use 10-15 ml of fixative per 1-2 fish in a sealable container. The volume of fixative should be at least 10 times the volume of the specimen

This is a sample clip. If you're new to JoVE sign up and start your free trial today to watch the full video! If your institution has an existing subscription, log in or sign up to access this video The Zebrafish International Resource Center is a non-profit organization. The prices we charge for products and services are to offset some of the actual costs we incur In cancer biology, a tumor begins from a single cell within a group of precancerous cells that share genetic mutations. Kaufman et al. used a zebrafish melanoma model to visualize cancer initiation (see the Perspective by Boumahdi and Blanpain). They used a fluorescent reporter that specifically lit up neural crest progenitors that are only present during embryogenesis or during adult melanoma. (A) At 30 hpf, heg1 is expressed throughout the entire endocardium. (B) Later, at 48 hpf, heg1 expression becomes restricted to high fluid shear stress regions of the endocardium including the atrioventricular canal (AVC).(C) The strong expression of heg1 at the AVC persists at 72 hpf.(D) Expression of heg1 was reduced in morphant embryos lacking blood-flow [troponin T type 2a (tnnt2a)] at 48 hpf * MESAB (MS-222 or Tricaine) - This is a very common fish anesthesia. It's chemical name is MESAB, 3-amino-benzoic acid ethyl ester and it can be ordered from Sigma (1-800-325-3010). One needs to be careful with MESAB. According to the Material Safety Data Sheet, it's potentially an irritant

ENU Mutagenesis Procedure With - zfin

  1. Store fixative at room temperature. 30 ml Ethanol (95%) 10 ml Formalin (Formaldehyde 37% solution containing 10-15% methanol, Sigma # F1635) 2 ml Glacial Acetic Acid. 58 ml Distilled Water. Euthanatize fish with an overdose of MS-222 (tricaine methane sulfonate). Carefully slit open the body cavity along the belly, trying not to disrupt.
  2. utes following cessation of opercular movement
  3. information about sequences are typically obtained from original publications or ZFIN, NCBI, and Ensemble websites. In addition to primer sequences, MacVector also provides calculations and predictions about the T m (melting temperature) of primers and the optimal annealing temperature for each primer set. Typically, we design primer set
  4. cells Article Zebrafish Microenvironment Elevates EMT and CSC-Like Phenotype of Engrafted Prostate Cancer Cells Lanpeng Chen 1, Maciej Boleslaw Olszewski 2,3, Marianna Kruithof-de Julio 4,5 and B. Ewa Snaar-Jagalska 1,* 1 Institute of Biology, Leiden University, 2311 EZ Leiden, Netherlands; l.chen@biology.leidenuniv.nl 2 Department of Molecular Biology, International Institute of Molecular.
  5. The tricaine anesthetization is the standard procedure in zebrafish experiments. After lesion, the whole sample preparation process does not use any anesthetic drugs. Before they were sacrificed, we used ice-cold water to immobilize them and quickly dissected the cerebellum out. , zebrafish-human gene ortholog data (ZFIN) , and zebrafish.
  6. Tricaine Methanesulfonate (MS-222) is a popular anesthetic agent used in aquatic species, and is intended for the temporary immobilization of fish, amphibians, and other aquatic, cold-blooded animals. It has long been recognized as a valuable tool for the proper handling of these animals during manua

ZFIN The Zebrafish Information Networ

ZFIN Cite Us . How to Mount Zebrafish in Methylcellulose. Using Methylcellulose is convenient for mounting live embryos for observation under the compound microscope. Methylcellulose is a plant extract that is dissolved in fish water to make a viscous solution. It puts a relatively loose hold on the embryo, so you can move the embryo to. Embryos may be anesthetized using 16 mg mL − 1 tricaine prior to imaging if desired (Westerfield, 1995). Note: Analysis of acridine orange staining in zebrafish embryos is complicated by autofluorescence and/or nonspecific uptake by the yolk, iridophores, hatching gland, and neuromasts ( Rodriguez & Driever, 1997 )

Zebrafish at 48, 78, and 100 hpf were mounted in 2% low-melting agarose without Tricaine. 20-30 s movies were recorded with 5 ms exposure. Light intensity and duration were kept to a minimum to avoid light-induced twitching For imaging, embryos were mounted in 0.7% low-melting-point agarose-×0.3 Danieau-tricaine) in glass bottom culture dishes (MatTek) with the dorsal aspect of the neural tube facing the coverslip E3 medium (for zebrafish embryos) 34.8 g NaCl. 1.6 g KCl. 5.8 g CaCl 2 ·2H 2 O . 9.78 g MgCl 2 ·6H 2 O . To prepare a 60X stock, dissolve the ingredients in H 2 O, to a final volume of 2 L. Adjust the pH to 7.2 with NaOH. Autoclave. To prepare 1X medium, dilute 16.5 mL of the 60X stock to 1 L. Add 100 µL of 1% methylene blue (Sigma-Aldrich) Pentair Aquatic Eco-Systems. During this COVID-19 pandemic, Pentair global operations and supply teams are working diligently to help ensure our valued customers are getting the best possible service and delivery during this time. Despite our efforts, shipping delays may occur Fish were anesthetized with 0.004% Tricaine. Metamorphic and post-metamorphic fish were measured and staged according to [ 26 ], treated with 1mg/ml (-) Epinephrine to cause pigment granule aggregation and photographed with a Leica M205 FA stereomicroscope with a Leica DCF300 FX camera using the software LAS V4.1 to allow multifocus images

ZFIN: Zebrafish Book: Content

  1. After the desired exposure to diverse ligands, larvae were euthanized with Tricaine (MS-222, 300 µg/mL) and placed in Trizol for RNA isolation. The same experimental layout was performed 5 times, and samples were subjected to microarray hybridization and analysis. Overall design: Microarray experiments were performed as single-color.
  2. Store fixative at room temperature. 30 ml Ethanol (95%) 10 ml Formalin (Formaldehyde 37% solution containing 10-15% methanol, Sigma # F1635) 2 ml Glacial Acetic Acid. 58 ml Distilled Water. Euthanatize fish with an overdose of MS-222 (tricaine methane sulfonate). Carefully slit open the body cavity along the belly, trying not to disrupt.
  3. The zebrafish mutant strain clo m39 (RRID:ZFIN_ZDB-ALT-980203-381), the fgf20a enhancer-trap line HGn21A (Shibata et al., 2016), the BAC Tg(il1b:egfp) line, and Tg(hsp70l:il1b) were used. The clo m39 mutant was genotyped as previously described (Hasegawa et al., 2015). Zebrafish were subject to heat-shock induction at 38°C for 1 hr in a small.
  4. The scl PAC clone (BUSM-129I22) was purchased from the Chris T. Amemiya laboratory via ZFIN (www.zfin.org) For time-lapse confocal microscopy, embryos were first anaesthetized in 0.02% tricaine in embryo water, then mounted in 0.5% low-melting agarose on a 35-mm plastic Petri dish (Iwaki), and covered with embryo water containing 0.02%.

Tricaine methanesulfonate (MS-222) on the spermatic

  1. MESAB/tricaine diluted in fish water (recipe in The Zebrafish Book) 3. DRY FISH: Once anesthetized, gently blot fish dry on paper towel, paying special attention to dry ventral side. Water activates sperm so it is important to thoroughly dry around the cloacae. Position fish on sponge holder, ventral side up (Fig. 2)
  2. Microfibril-associated glycoprotein 4 (MFAP4) is an extracellular matrix protein belonging to the fibrinogen-related protein superfamily. MFAP4 is produced by vascular smooth muscle cells and is.
  3. Introduction. The ability to predict threats or rewards and to shape behavior in response to a changing environment is essential for survival (Skinner, 1984).Operant learning requires a continuous monitoring of the animal's own actions, a real-time estimation of its consequences, and an update of the relationship between the behavior and its outcome (Skinner, 1981)
  4. Left-Right Identity of the dHb Influences Fear Responses. Directional asymmetry of the dHb can be altered in predictable ways in zebrafish. Ablation of the parapineal, an accessory organ to the pineal, results in right isomerization of the dHb, in which both nuclei develop symmetrically with right identity [ 12, 13 ]
  5. obenzoate methanesulfonate salt) added to external solution; can also add 100-200 nM TTX to external solution to reduce muscle twitching. Stock Solution: 400 mg tricaine powder 97.9 ml DD water ~2.1 ml 1 M Tris (pH 9). Adjust pH to ~7. Store at -20˚ C. Working Solution: 0.5 ml tricaine stock solutio
  6. Zebrafish (Danio rerio) are being used in increasing numbers within molecular biology, developmental biology, neurobiology, genetics, cancer research and drug discovery, due to the low costs of maintaining them, their short generation interval, the transparency of the embryos and the ability to manipulate the genome.There are frequent meetings about the care and use of zebrafish

Endothelial cell sprouting is a fundamental process of physiological and pathological blood vessel growth. Attracted by growth factors such as vascular endothelial growth factor-A (VEGF-A) secreted from hypoxic tissues, endothelial cells (ECs) break out of the quiescent vessel wall to form new vessel branches (Ferrara et al., 2003; Koch and Claesson-Welsh, 2012) Fish were washed in ocean salt water (0.3g/L aquarium salt) and then transferred into ocean salt water and incubated for 10 minutes in the dark at room temperature. After incubation zebrafish were euthanized with 0.4% Tricaine (Sigma, MO), 20mM Tris pH 9.0 and mounted in 3% methyl-cellulose 2.0 Effective Date: 6/2016 3.0 Purpose: The purpose of this policy is to outline acceptable methods of euthanasia for laboratory animals at the University of Oregon. 4.0 Scope: This policy applies to all research investigators and animal care staff at the University of Oregon whose IACUC approved Animal Use Protocols (AUP) require euthanasia of animal subjects A loss of Ccm proteins in zebrafish or mice results in higher levels of klf2 mRNA expression, suggesting that the physiological role of Ccm proteins is to modulate klf2 expression levels in response to blood-flow (Renz et al., 2015; Zhou et al., 2015, 2016).Given that levels of heg1 mRNA expression are affected by blood-flow, we explored whether upregulation of Heg1 or Krit1 would have an.

Anesthetize embryos by transferring a dechorionated embryo into 0.16 mg/ml Tricaine (3-amino benzoic acidethylester, Sigma-Aldrich) solution (40μl of 4 mg/ml stock solution in 1 ml of E3 medium 1X). Note 1: Transfer embryos with a glass pipette or a cut-tip (P1000) to minimize the risk of damaging the embryos Adult fish were 6-10 months old and had a body length of 25-32 mm. Fish were anaesthetized using Tricaine (Sigma). A canula (30 gauge, outer diameter 300 μm) was pushed through a nostril ∼6-8 mm deep along the rostrocaudal body axis, through the olfactory bulb until reaching the caudal part of the telencephalon Animals and husbandry. Eighteen male zebrafish (AB/wt, date of fertilisation: 7 th of February 2013, weight: mean ± SD; 0.25±0.03 g) were used for this experiment. Two fish from the control group and one fish from the tricaine group had to be excluded due to methodological problems, leaving eight fish in the tricaine group and seven fish in the control group

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MS222 (tricaine methanesulfonate, Ethyl 3-aminobenzoate) • sulfonated analog of benzocaine (makes it more hydrophilic) • inhibits sodium ion channels • acts systemically in fish • analgesic, sedative and paralytic activity shown to block motor and sensory neurons in Xenopus explants. Ramlochansingh et al. PLoS One. 2014; 9(7): e101606 Most cases of Usher syndrome type II (USH2) are due to mutations in the USH2A gene. There are no effective treatments or ideal animal models for this disease, and the pathological mechanisms of USH2 caused by USH2A mutations are still unknown. Here, we constructed a ush2a knockout (ush2a−/−) zebrafish model using TALEN technology to investigate the molecular pathology of USH2

Sacrifice donor zebrafish in 1.6 mg/ml tricaine methanesulfonate (MS222) for 10 min or until no operculum movement is evident. Place donor fish on an absorbent paper towel and excise the dorsal muscle using a clean razor blade. The cut should be made near the anus at a 45° angle to maximize tissue collection (as noted in Figure 1A). Place. in 0.003% tricaine (3-amino benzoic acid ethyl ester; Sigma Chemical Co.) at pH 7 (Westerfield, 1994). Re- peated anesthesia and rinsing appears to slightly but significantly retard subsequent development (see leg- end to Fig. 2). Nomarski Optics A compound microscope equipped with Nomarski dif Before imaging at 3 dpf, tricaine was removed and embryos were allowed to recover in Evans solution (134 m m NaCl, 2.9 m m KCl, 2.1 m m CaCl 2, 1.2 m m MgCl 2, 10 m m glucose, 10 m m HEPES) for 10 min and then treated with 200 m m N-benzyl-p-toluene sulphonamide to inhibit contraction for 5 min. KCl, 100 μ m, was added to initiate muscle.

2. Bring to fish facility: ice bucket, Hanks, Tricaine, timer, tubes for collecting sperm. 3. Dilute Tricaine in fish water 3. Collect sperm 1. Prepare a clear vial of Hanks solution on ice. Measure ~50 μl per male to be squeezed. 2. Anesthetize 3 or 4 males at a time by immersion in tricaine. They can be lifted out of the anesthetic with a. Embryos aged between 24 hpf and 92 hpf were anesthetized with Tricaine(Sigma) and immobilized on a plate of 3% agarose. BrdU 100 mM (5 μl; Sigma)was injected into the brain ventricle. Injected embryos were allowed to develop until 96 hpf when they were fixed with 4% paraformaldehyde (Sigma) Zebrafish were sedated in tricaine as described by others (Westerfield, 2007), but with some differences in drug preparation. A 40μg/L solution of tricaine (tricaine, methanesulfonate, or MS-222, Sigma), dissolved in fish water (RO water with 0.2g/L Instant Ocean and 0.1g/L sodium bicarbonate) o Embryos should be euthanized in a solution of 250 mg/L Tricaine. Tricaine is available through Fisher (AC11800-0050). Acoustic StartleTest Materials Timer or stopwatch Petri dish (10 cm) (either plastic or glass) Beaker Plastic transfer pipettes Egg wate Zebrafish Information Network (ZFIN), the model organism database for zebrafish (Bradford et al., 2011), contains ~9900 mutant lines of which 62% of these chemically induced lines remain uncloned. Presumably, both the actual number and percentage of uncloned mutants is even higher than this figure because many researchers wish to clone mutants.

ZFIN All Figures, Shwartz et alZFIN All Figures, Bussmann et al

Zebrafish has emerged as an important animal model to study human diseases, especially cancer. Along with the robust transgenic and genome editing technologies applied in zebrafish modeling, the ease of maintenance, high-yield productivity, and powerful live imaging altogether make the zebrafish a valuable model system to study metastasis and cellular and molecular bases underlying this. Original Article Valvular and Mural Endocardiosis in Aging Zebrafish (Danio rerio) T. K. Cooper1 and J. M. Spitsbergen2 Abstract Endocardiosis or myxomatous degeneration of the cardiac valves is a well-described age-related change in humans and dogs

Recipes - Protocols - ZFI

Introduction and Objectives The zinc-finger transcription factor Krϋppel-like factor 2 (KLF2) transduces blood flow into molecular signals responsible for a wide range of responses within the vasculature. KLF2 maintains a healthy, quiescent endothelial phenotype. Previous studies report a range of phenotypes following morpholino antisense oligonucleotide-induced klf2a knockdown in zebrafish Intriguingly, genetic lineage tracing has shown that mature osteoblasts do not contribute to bone repair in mammals (Park et al., 2012), and dedifferentiation of mature osteoblasts is not considered to occur in mammalian bone biology [with the possible exception of pediatric osteosarcomas (Pereira et al., 2009)].Instead, during mammalian fracture healing, new osteoblasts appear to be supplied.

Methyl Cellulose Mounting - Protocols - ZFI

pxr exon 2 (ZFIN: ZDB-GENE-030903-3) was queried for putative targets using the web tool CHOPCHOP (Howe et al., 2013; Montague et al., 2014) and the zebrafish GRCv10 genome. From this, we selected 3 targets opting for sequences that contained a G nucleotide within the first 3 nucleotides of the target se-quence and zero predicted off. Briefly, adult zebrafish at appropriate stages were anesthetized in tricaine (0.016%) for 5 minutes, placed ventral side up into a sponge. The MX700 transducer was placed above the zebrafish to provide a sagittal imaging plane for the heart. Image quantification was performed using the VevoLAB workstation

After the bath treatment, all embryos were collected in 1.5 ml taining 0.16 mg/ml tricaine. Each embryo was inoculated with 10 bacteria Eppendorf tubes containing 500 ml TRIzol reagent (Invitrogen) and into the yolk sac following a microinjection procedure Evaluation of rapid cooling and tricaine methanesulfonate (MS222) as methods of euthanasia in zebrafish (Danio rerio). Journal of the American Association for Laboratory Animal Science, 48, 785-789 At 2 or 3 dpf the PTU treated embryos were anesthetized in 160ug/ml tricaine methane sulfonate and mounted in 1.2% low-melting agarose/160ug/ml tricaine methane sulfonate. ZFIN Direct Data.

For light-sheet microscopy, zebrafish embryos were anesthetized with 0.003% tricaine, embedded in 1% low melting agarose with tricaine and mounted in Zeiss light-sheet Z.1 capillary specimen holder, vertically immersed in imaging chamber containing water with tricaine In the Zebrafish Model Organism Database (ZFIN), the mutant allele designation for this zebrafish line is brca2 hg5. Histological Analyses. Zebrafish were euthanized with tricaine methanesulfonate and fixed in 4% paraformaldehyde at 4 °C for a minimum of 24 h before being transferred to 70% ethanol Adult wildtype and erbb3b −/− zebrafish (∼8 months old) were killed with Tricaine, fin tissue was collected for genotyping, and the trunk region was dissected and placed in fixative: 2% glutaraldehyde/4% PFA in 0.1 m sodium cacodylate. Adult zebrafish trunk samples were prepared for electron microscopy using microwave fixation (Panasonic. For confocal imaging, fish were anesthetized with 0.02% tricaine and mounted in 1.2% low melt agarose (Promega, V2111). Fluorescence was imaged with a Zeiss LSM 510 confocal Page 6 of 42 John Wiley & Sons, Inc. Developmental Neurobiolog using 3-aminobenzoic acid ester (Tricaine), immersed in 0.8% low-melting point agarose, and mounted on their sides in glass-bottom, 35 mm Petri dishes (Electron Microscopy Sciences). Images were captured using either a 40 (numerical aperture 1.2) or a 63 (numerical aper-ture 1.2) water-immersion objective mounted on a motorized AxioOb

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Using tricaine to anesthetize embryos. In our hands, tricaine has occasionally induced improper staining patterns in embryos; thus, it is best to use ice to anesthetize embryos prior to imaging. The inability to use chemical anesthetics also limits the embryonic stages that can be feasibly imaged Zebrafish larvae were either provided with or deprived of food according to the feeding protocol described earlier. 7 dpf larvae from each group were pooled in sets of 24 individuals, anesthetized with 0.016% tricaine in Danieau's medium, and lysated with a tissue homogenizer. Cortisol was extracted following a previously described protoco

Morpholinos blocking tp53, 27 (Zfin tp53 MO-4) and has2 28 (Zfin has2 MO-1) are previously described. All morpholinos were supplied by GeneTools and diluted to a 1 mM stock. Working concentrations were as follows: hapln1a 500 nM or 250 nM, has2 250 nM, combinatorial has2/hapln1a 250 nM each, tp53 250 nM Zebrafish Information Network (ZFIN) clone CB330 (19). Alignment of the completely sequenced clone CG385 with AE2 sequences Female X. laevis anesthetized with 0.17% tricaine were subjected to partial ovariectomy in accordance with protocols approved by the Institutional Animal Care and Use Committee o

NOTE: Dose of tricaine anesthesia will depend on age and size of recipient zebrafish. 3. Place anesthetized recipient zebrafish on a damp paper towel or sponge, with the left side facing up. 4. Insert the syringe needle into the latero-dorsal musculature (refer to Figure 1A). Ensure that injections are performed at a 45° angle. Inject Adult fish were anesthetized in 0.04% Tricaine (Sigma) in fish water prior to stab lesion. Stab lesions were performed as described in Kroehne et al. (2011). In brief, a 30 gauge cannula was inserted through a single nostril along the rostrocaudal brain-axis, into the olfactory bulb and finally the caudal aspect of the telencephalic hemisphere qrfp (ZFIN gene name si:ch211-185o22.2) mutant.. ZFN binding sites were 5′-ACAGTCTTC-3′ and 5′-AGCACCAAC-3′. qrfp mutant line i4 contains a 4 bp insertion (TTCT, after nucleotide 51 of the open reading frame). The mutation results in a change in reading frame after amino acid (aa) 18 and a premature stop codon after aa 42, compared with 168 aa for the WT protein come from the ZFIN database, where in situ hybridiza-tion staining experiments from high-throughput expression screens and individual publications have been annotated in detail using the AO system. ZFIN provides two views of the anatomical information on gene expression: first, anatomical terms for individual genes on ZFIN are listed on the gen The ruthenium-based photosensitizer (PS) TLD1433 has completed a phase I clinical trial for photodynamic therapy (PDT) treatment of bladder cancer. Here, we investigated a possible repurposing of this drug for treatment of conjunctival melanoma (CM). CM is a rare but often deadly ocular cancer. The efficacy of TLD1433 was tested on several cell lines from CM (CRMM1, CRMM2 and CM2005), uveal. Time-lapse micrography was performed on larvae anaesthetised in tricaine and mounted in agar as described in The Zebrafish Book (Westerfield, 1995). Images were taken on a Zeiss Axioplan microscope equipped with a DAGE MTI camera controlled by NIH Image, at a rate of one frame every minute for primordium migration, or every 1.3 minutes for the.