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15 protocols using u 100 insulin syringe

1

Adult Zebrafish Brain Injury Protocol

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Adult brain injury was performed as described previously (Marz et al., 2011 (link)). Adult zebrafish were anesthetized with Tricaine (Sigma) and a BD U-100 insulin syringe (BD bioscience) was inserted vertically through the skull into the telencephalic region. Then brain damaged fish were transferred into a tank with fresh fish water and fixed in 4% paraformaldehyde at 4-day post injury.
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2

Topical Remineralization Agents Comparison

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Both fluoride varnish and nano-HAP paste were applied using 1 cm 3 BD™ U-100 insulin syringe (BD, Franklin Lakes, New Jersey, USA) to standardize the amount of agent applied on the lesion surface. A quantity of 0.02 cm 3 of both agents was sufficient to cover the lesion surface of the specimens. 6 In Group II, fluoride varnish was applied and airdried for 30 seconds. The specimens were stored in the remineralizing solution. After 4 hours, the varnish was removed with a scalpel blade gently to avoid touching the lesion surface. The surface of every specimen was washed using a syringe containing the remineralization solution: 1.5 mM CaCl 2 , 0.9 mM NaH 2 PO 4 , and 0.15 M KCl at pH of 7. 14 In Group III, according to the manufacturer's instructions, nano-HAP paste was applied on the specimen followed by friction of 10 seconds using a microbrush, then it was left in contact with the lesion surface for 4 minutes and removed with deionized water. This was done once a day for 2 days separated by one pH cycle.
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3

Streptozotocin-Induced Diabetic Embryopathy

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We8 , 10 (link), 25 (link)–29 (link) and others30 (link)–32 (link) have used a rodent model of Streptozotocin (STZ)-induced diabetes to study diabetic embryopathy. Briefly, ten-week-old WT female mice were intravenously injected daily with 75 mg/kg Streptozotocin for two days to induce diabetes. Streptozotocin from Sigma (St. Louis, MO) was dissolved in 0.1 M citrate buffer (pH 4.5). We used a U-100 insulin syringe (Becton Dickinson, Franklin Lakes, NJ) with 281/2-G needles for injections. Approximately 140 μl of STZ solution was injected per mouse. Diabetes was defined as a 12-hour fasting blood glucose level of ≥ 16.7 mM. Male and female mice were paired at 3:00 P.M., and day 0.5 (E0.5) of pregnancy was established at noon on the day that a vaginal plug was present. Embryos were harvested at embryonic day (E) E8.5 (2:00 PM at E8.5) for biochemical and molecular analyses.
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4

Culturing Murine Bone Marrow Cells for AZT Exposure

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Bone marrow cells from two to four C57BL/6J mice/genotype were rinsed from the femur and tibia using a disposable LO-DOSE 1/2 cc U-100 Insulin Syringe with a permanently attached 28G 1/2 needle (Becton Dickinson, Franklin Lakes, NJ). The material was resuspended in RPMI media (ATCC, Manassas, VA) containing 10% fetal bovine serum (ATCC) and grown in T-75 Falcon flasks (BD Falcon, Bedford, MA) until they reached 95% confluency. To remove cells from flasks the monolayer was washed with Dulbecco’s phosphate buffered saline (DPBS, Gibco Invitrogen, Carlsbad, CA) without Ca and Mg, twice and incubated with 0.05% Trypsin (Gibco Invitrogen) for 4 minutes at 37°C. After neutralizing with an equal volume of media and centrifuging at 2,000 rpm for 4 minutes, cells were resuspended and distributed into 4-well culture slides (BD Falcon). Following a 4-hr attachment period, the media was replaced with media containing 0, 10, or 100 μM AZT (Sigma-Aldrich Co, St. Louis, MO) and cells were incubated for 24 hr. To prepare a stock solution, AZT was dissolved in DPBS and the final concentration was calculated using an extinction coefficient of 11,500 m2/mol at an absorbance wavelength of 266 nm.
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5

Intradermal S. aureus Infection Model in Mice

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All procedures were approved by the Regional Commission of Freiburg, Baden - Württemberg. Approximately 107colony forming units (CFUs) S. aureus in 10 µl PBS were intradermally (i.d.) injected (30 gauge needle, U-100 insulin syringe (BD)) into the ear pinna of anaesthetized mice. In some experiments, heat fixed (hf) bacteria were used for i.d. inoculation (108 hf bacteria in 10 µl PBS per ear pinna). For re-infection experiments either 107 CFU S. aureus in 10 µl PBS or 108 hf bacteria in 10 µl PBS were i.d. injected into the same ear pinna 3 weeks (if not indicated otherwise) after the first infection. If not indicated otherwise groups of 4–5 mice were used per experiment followed, by at least one repetition to confirm the results. Lesion morphology was documented by digital photographs (Canon PowerShot A650 IS) of mice ears and analyzed by the software program Adobe Photoshop CS6.
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6

Neuronal Activity Modulation in Temporal Muscle

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When the ongoing (unstimulated) neuronal activity of a unit was visibly stable, it was recorded for a control period of 30 min without any treatment (baseline activity). Subsequently, saline (0.9 % NaCl, 50 μl) was injected with a U-100 insulin syringe (BD, Franklin Lakes, NJ, USA) deep into the temporal muscle close to the crista temporalis within one minute. After another 30 min of recording, 50 μl of 10 mM α,β-meATP (α,β-methylene adenosine 5′-triphosphate disodium salt hydrate, Sigma-Aldrich, Taufkirchen, Germany) was injected into the temporal muscle and the recording continued for 60 min. After another 10 min without further treatment (new baseline), in most of the experiments 60 μl of 2 % lidocaine (Xylocain, Astra Zeneca, Wedel, Germany) was injected into the ipsilateral medial (semispinalis and rectus capitis) neck muscles followed by a recording period of 20 min. Next the temporal muscle was injected with 60 μl lidocaine and the activity recorded for another 20 min. Finally, 60 μl of lidocaine was applied onto the exposed cranial dura mater followed by a final recording period of 20 min. In some experiments 60 μl of lidocaine was injected only into the ipsilateral temporal muscle immediately followed by 50 μl of α,β-meATP, then the recording was continued for 60 min.
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7

Pathogen Injection and Lymph Node Sampling

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C57BL/6J mice (4 and 22 months old) were purchased from Harlan NIA as part of the age-controlled NIH mouse colony program. All animal procedures were carried out according to a protocol approved by the Institutional Animal Care of Albert Einstein College of Medicine. In some experiments, mice were injected in the footpad of both legs with tree different pathogens: a nonvirulent strain of C. neoformans (H99), stained with 1% Uvitex 2B, with M. smegmatis expressing GFP, and with S. aureus (Wood strain without protein A) BioParticles®, Alexa Fluor® 488 conjugate (Molecular Probes®, San Diego, CA, USA).
An amount between 1 × 107 was injected in each leg with a 3/10 mL BD Lo-Dose™ (Franklin Lakes, NJ, USA) U-100 insulin syringe with 29 G × 1/2 in. BD Ultra-Fine™ (Franklin Lakes, NJ, USA) IV permanently attached needle. After different time points (5 min, 20 min, 1 h, and 2 h), the popliteal lymph node (PLN) and collector running from the injection site to the PLN were collected. The derma of the injection site was collected after 5 min as well.
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8

Tamoxifen-Induced Lineage Tracing in Mice

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Tamoxifen (TAM) was administered by subcutaneous (s.c.) injections (BD Micro-Fine 0.5 mL; 29G; U-100 Insulin Syringe; REF 324824) up to three times between postnatal (P) days 4 and 14 with a maximum of one dose per day. Mice were induced with 100 mg/kg body weight TAM (Sigma-Aldrich; T5648) dissolved in corn oil (Sigma-Aldrich; C8267). As negative controls, corn oil-induced PDGFRβ-P2A-CreERT2-tdTomato mice were used.
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9

Tongue Tumor Xenograft Model

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Nude mice (8 weeks of age) or WT and TNCKO (C57Bl/6J) mice (bred in house) were grafted in the first third part of the tongue. In particular, 3 × 106 OSCC13 cells in 10 µl PBS were injected using a U-100 insulin syringe (BD Micro-Fine) in C57Bl/6J mice. Some 1 × 106 cells OSCC13-LUC (luciferase expression vector expressing cells) were similarly grafted. Tumors were visible 2 weeks upon engraftment and mice were sacrificed for analysis at 3 weeks (bioluminescence experiment), 2 and 4 weeks (NIR) or 4 weeks (IR). For irradiation analysis, a 2 Gy unique dose of photons was delivered two weeks after tumor cell engraftment (3 × 106 cells). Mice were sacrificed two weeks later and tissue was analyzed by staining. For bioluminescence detection, a RediJect D-Luciferin Ultra Bioluminescent Substrate (Perkinelmer 770505) solution at 30 mg/ml was injected intraperitoneally 7 min before imaging. Images were acquired for 5 min using a live imager (NightOwl, Berthold). All mice were housed and handled according to the guidelines of INSERM and the ethical committee of Alsace, France (CREMEAS) (Directive 2010/63/EU, 01386.02, C-67-482-033 on the protection of animals used for scientific purposes).
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10

Insect Infection Assay Protocol

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A U-100 insulin syringe equipped with a 29-gauge needle (BD) was used to inject 10 μL aliquots of a 1 × 106 cfu mL−1 bacterial inoculum in PBS or 10 mM MgCl2 into the haemocoel of each G. mellonella larvae via the last left proleg. Before and after injection, the area was cleaned using an alcohol swab. After injection, G. mellonella larvae were incubated in plastic containers and kept at room temperature in the dark. The number of dead larvae were scored daily for 72 h. Larvae were considered dead when they displayed no movement in response to a stab to the head. The crude cell-free supernatant (derived by centrifuging and filtering (0.2 μM)) from an overnight (O/N) broth culture of the bacterial strains was also used to assess toxicity.
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