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29 gauge needle

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The 29-gauge needle is a medical instrument designed for precise and minimally invasive procedures. It is a small-diameter needle typically used for administering injections or collecting samples. The core function of the 29-gauge needle is to provide a gentle and controlled means of access to the body, allowing healthcare professionals to perform their duties effectively.

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12 protocols using 29 gauge needle

1

Intracardiac Xenograft Model of Metastasis

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As previously described,40 4‐week‐old male nude mice were anesthetized with tribromoethanol. U2OS‐luc cells (1 × 106) were resuspended in PBS and inoculated into their left ventricles using tuberculin syringes with 29‐gauge needles (BD bioscience). When the needle was implanted, blood jets were observed as a criterion for entering the left ventricle, and implantation was completed within 10 seconds. Three days later, Tan I (10 or 20 mg/kg/d) was intraperitoneally administrated to the mice for 32 days. On days 0, 12 and 24 post‐Tan I treatment, tumour metastasis was monitored using bioluminescence imaging after injection of D‐luciferin (Gold Biotechnology). Imaging data were analysed with the Living Image Software (Calliper Life Sciences). Survival curves of mice were also recorded.
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2

Retroviral Transduction of VGLL3 in Cardiac Myofibroblasts

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Plat-E cells were transfected with EGFP/pMXs-puro or EGFP-VGLL3/pMXs-puro using PEI-Max. Forty-eight h after transfection, the supernatant containing the retrovirus was concentrated by centrifugation (8000 × g) at 4 °C for 16 h, and precipitated retrovirus was suspended with PBS containing 4 μg/mL polybrene. For in vivo retroviral transduction of VGLL3 to cardiac myofibroblasts, this retroviral suspension (20 μL/heart) was administrated intramyocardially just below the ligation site immediately after the ligation of the coronary artery, using 29-gauge needles (BD Biosciences). Three days after administration, mouse hearts were fixed in 4% PFA/0.1 M PB for 3.5 h. The intracellular localisation of EGFP-VGLL3 or EGFP in cardiac myofibroblasts was assessed using immunohistochemistry.
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3

Comparative Assessment of AAV Delivery Routes

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A standardized dose of 1×1011 vg was delivered to each mouse using four routes of administration: intravenous (IV), intramuscular (IM), intraperitoneal (IP), or intranasal (IN). Mice were warmed under a heat lamp and restrained in a tail vein injection device to administer IV injections. Vectors were diluted in PBS to 100 μl and injected with a 29-gauge needle (BD Biosciences, California, USA). To administer IM injections, mice were restrained in a 50 mL tube with the end removed. Fur was shaved around the gastrocnemius muscle and vectors were injected in a 40 μl volume using a 27-gauge tuberculin syringe. Mice were restrained by the scruff and injected IP with a 100 μl dose in the intraperitoneal cavity using a 27-gague tuberculin syringe. Mice were lightly anesthetized with isoflurane for IN administration and restrained as previously described (24 ). The vector was diluted in PBS to 80 μl and delivered in two rounds of administration (40 μl each) separated by a recovery period of 20 minutes.
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4

Cardiac Injection of miR-22 and Exosomes

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With the chest open, synthetic oligonucleotides pretreated with 15 µl Dharmafect Duo (Thermo) were injected into the myocardium through an insulin syringe with a 29-gauge needle (BD). The dosages used per mouse were 80 ng miR-22 mimic, 1 nmol Mecp2 siRNAs On-TARGETplus SMART pool (a pool of four target-specific 20–25 nt siRNAs; Thermo). One µg exosomes (Exonon-IPC, ExoIPC, ExoIPC+miR-22 Inhibitor) were injected along the border between infarct zone and normal myocardium after LAD.
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5

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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6

Comparative Assessment of AAV Delivery Routes

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A standardized dose of 1×1011 vg was delivered to each mouse using four routes of administration: intravenous (IV), intramuscular (IM), intraperitoneal (IP), or intranasal (IN). Mice were warmed under a heat lamp and restrained in a tail vein injection device to administer IV injections. Vectors were diluted in PBS to 100 μl and injected with a 29-gauge needle (BD Biosciences, California, USA). To administer IM injections, mice were restrained in a 50 mL tube with the end removed. Fur was shaved around the gastrocnemius muscle and vectors were injected in a 40 μl volume using a 27-gauge tuberculin syringe. Mice were restrained by the scruff and injected IP with a 100 μl dose in the intraperitoneal cavity using a 27-gague tuberculin syringe. Mice were lightly anesthetized with isoflurane for IN administration and restrained as previously described (24 ). The vector was diluted in PBS to 80 μl and delivered in two rounds of administration (40 μl each) separated by a recovery period of 20 minutes.
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7

In vivo Implantation of Stem Cell Constructs

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For in vivo implantation, cells were divided into six groups, 1 × 106 mouse dermal cells alone used as a control group (n =8), and 5 × 105 P1-HFSCs mixed with 5 × 105 mouse dermal cells, which were used in five different experimental groups (n =8 per group): HFSCs, LbL-coated HFSCs (LbL-HFSCs), HFSCs + TGF-β2 (10 pM), LbL-coated HFSCs loaded with TGF-β2 [LbL(TGF-β2)-HFSCs], and 50% LbL-HFSCs + 50% LbL (TGF-β2)-HFSCs (50% + 50%). Athymic nude mice were anesthetized with pentobarbital sodium (1.3 mg/kg body weight). Next, 1 × 106 cells from each group were injected subcutaneously into the dorsal side in a total volume of 50 µL PBS using a 29-gauge needle (BD Biosciences). After 3 wk, the grafted specimens were examined under a stereomicroscope (MVX10, Olympus) and photographed. The grafted regions were harvested, fixed in 4% paraformaldehyde, and embedded in paraffin. Subsequently, 4-µm thick serial sections were stained with hematoxylin and eosin (H&E). Representative areas for each group were selected and photographed for further analysis.
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8

Leishmania infantum Infection in Hamsters

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Leishmania infantum (MCAN/BR/00/BA262) promastigotes isolated from a naturally infected dog (Bahia State, Brazil) were cultured in Schneider’s medium (LGC, Brazil) supplemented with 10% of inactivated FBS (fetal bovine serum) (Gibco, USA), 2 mM L-glutamine, 100 IU/ml penicillin, 1% streptomycin (Gibco, USA).
Fifteen days after the last immunization, hamsters were inoculated by intradermal route, in the left ear with 105 stationary phase promastigotes plus 0.5 pair of sonicated salivary glands (SGH) from female Lutzomyia longipalpis, using a 29-gauge needle (BD Ultra-Fine) in 20uL of saline. Salivary glands were dissected from 5- to 7-day-old females and stored in endotoxin-free PBS at −70°C. Salivary glands were sonicated (Sonifer 450 homogenizer, Branson, Danbury, Connecticut), and afterwards centrifuged at 12,000 g for 5 minutes. Supernatant was collected and used immediately.
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9

Immunization and Parasite Burden Analysis in Hamsters

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Six to eight-week-old female hamsters were immunized with 106 total stationary-phase LmexCen−/− parasites by intradermal injection in the left ear in 10 μl PBS using a 29-gauge needle (BD Ultra-Fine). In addition, the age-matched control group of animals were infected with 106 total stationary-phase L. mexicana wild-type (LmexWT) promastigotes. Lesion size was monitored weekly by measuring the diameter of the ear lesion using a direct reading vernier calliper. Parasite burdens in the ear, draining lymph node (dLN), spleen, liver and bone marrow tissues were determined by limiting dilution assay27 (link). Ear tissues were treated with Liberase enzyme (Sigma-Aldrich) (0.15 mg/ml) in DMEM mediun at 37 °C. After 90 min of incubation tissues were grinded to make single cell suspensison using BD medimachine system. Cells were washed two times with M199 medium. Lymph nodes, spleen, liver and bone marrow were harvested and homogenized with a cell strainer in 2 ml of M199 medium supplemented with 10% FBS and 1% Penicillin/Streptomycin. To determine parasite burden cell suspension from each organs were serially diluted in 96 well tissue culture plates. After 12 days of incubation at 26 °C, plates were examined with a inverted microscope and parasite burden were calculated for each organ.
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10

SDS-PAGE Analysis of Protein Samples

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Cells were subjected to three washes with PBS and subsequently lysed using SDS sample buffer (50 mM Tris-HCl, pH 6.8, 2% SDS, 100 mM dithiothreitol, 10% glycerol, and 0.025% bromophenol blue). High-molecular-weight DNA was sheared by passing the lysate through a 29-gauge needle (326,666, BD Biosciences). After centrifugation at 15,000 rpm for 5 min and heat treatment at 55 °C for 20 min, the samples were electrophoresed. Detection was performed using horseradish peroxidase (HRP)-conjugated secondary mouse or rabbit antibodies and chemiluminescence HRP substrate (WBKLS0100, Millipore-Sigma), and the signals were captured using an image analyzer (LAS500, GE Healthcare, Chicago, IL, USA).
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