Tumours were established by ovarian intrabursal (IB) implantation of ID8 cells as described previously [54 (link)]. Mice were anesthetised in an induction chamber using 3% isofluorane in 2 L/min oxygen and then maintained at 2% isofluorane in 1 L/min oxygen using a rodent facemask. An incision was made at the mid-dorsal region of the skin and the peritoneal membrane was excised at the latero-dorsal point above the location of the right ovary. The ovarian fat pad was externalised and stabilised with a serrefine clamp. Either 1 × 106 wild-type or pROSA-iRFP720 ID8 cells were loaded into a Hamilton microliter syringe (Sigma-Aldrich, St Louis, MO, USA) and injected underneath the ovarian bursa using a dissecting microscope for guidance. The skin was closed using Michel suture clips. Mice were monitored for recovery, and suture clips were removed 7 days post-surgery. Mice were fed an SF-AIN-93M rodent diet to reduce intrinsic autofluorescence. Mice were monitored weekly for weight and circumference, and humane endpoints were determined by a body abdominal circumference of >100 mm and general wellbeing (lack of responsiveness, hunched posture, ruffled fur, and eyes squinted). Once the endpoint was reached, mice were humanely sacrificed by CO2 asphyxiation, and blood and tissues were harvested for fluorescence imaging, ELISA, flow cytometry, and formalin fixed for immunofluorescence.
Hamilton microliter syringe
Manufactured by Merck Group
Sourced in United States
The Hamilton microliter syringe is a precision liquid handling instrument designed for accurate and reproducible fluid transfers. It features a borosilicate glass barrel and a tightly fitted plunger for precise volume control. The syringe is available in a range of sizes to accommodate various sample volumes.
Automatically generated - may contain errors
4 protocols using hamilton microliter syringe
1
Ovarian Tumor Induction via ID8 Cell Implantation
2
Intravitreal Amphiregulin Delivery in Mice
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A total of 5 µL AR-shRNA-AAV containing AAV-viral particles (1 × 1013 vg/mL for a total dose of 5 × 1010 vg) was intravitreally injected at baseline into the right eyes. For external amphiregulin, 20 ng murine recombinant amphiregulin was injected weekly after week 1. The intravitreal injections were performed under topical anesthesia using 0.5% proxymetacaine hydrochloride eye drops. The scleral entry for the injection needle was created by a 30-gauge needle puncture 1.5 mm posterior to the limbus. Using a Hamilton microsyringe (Hamilton Microliter syringe; Sigma-Aldrich, St. Louis, MO, USA), we delivered 5 µL of the solution into the right eyes through the scleral entry. The contralateral left eyes received an injection of 5 µL PBS.
3
Intravitreal Injection Protocol for Ophthalmic Drugs
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Everolimus and erlotinib, and MHY1485 (GlpBio Technology, Montclear, CA, USA) are insoluble in water. To keep a constant drug concentration during injection, suspensions were prepared by adding 5% polysorbate 80 (Sigma-Aldrich Co., St. Louis, MO, USA) to phosphate-buffered saline solution (PBS; Sigma-Aldrich Co.). All suspensions were immediately aliquoted and stored at −20°C. Intravitreal injections were performed under topical anesthesia using 0.5% proxymetacaine hydrochloride eye drops. The suspensions were thawed and shaken well before use. A 26-gauge needle (0.26 mm inner diameter) was used as a trocar needle to first make a vitreous entry port 2 mm posterior to the limbus. A 32-gauge Hamilton microsyringe (outer diameter: 0.235 mm, Hamilton Microliter syringe; Sigma-Aldrich Co.) delivered 5 µL of the solution into the eyes through the 26-gauge trocar needle.
All guinea pigs that received intravitreal injections of drugs into their right eyes received intravitreal injections of 5 µL of PBS with 5% polysorbate 80 into their left eyes. The guinea pigs in the negative control group and the NLIAE-only group received 5 µL of PBS with 5% polysorbate 80 once a week in both eyes.
All guinea pigs that received intravitreal injections of drugs into their right eyes received intravitreal injections of 5 µL of PBS with 5% polysorbate 80 into their left eyes. The guinea pigs in the negative control group and the NLIAE-only group received 5 µL of PBS with 5% polysorbate 80 once a week in both eyes.
4
Ovarian Cancer Tumor Induction via ID8 Cells
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Tumours were established by ovarian intrabursal (IB) implantation of pROSA-iRFP720 ID8 cells as described previously [22 (link),23 (link)]. Mice were anaesthetised in an induction chamber using 3% isofluorane in 1L/min oxygen and then maintained at 2% isofluorane in 0.3 L/min oxygen using a rodent facemask. An incision was made at the mid-dorsal region of the skin and the peritoneal membrane was excised at the latero-dorsal point above the location of the right ovary. The ovarian fat pad was externalised and stabilised with a serrefine clamp. 1 × 106pROSA-iRFP720 ID8 cells were loaded into a Hamilton microliter syringe (Sigma-Aldrich, St. Louis, MO, USA) and injected underneath the ovarian bursa using a dissecting microscope for guidance. The skin was closed using Michel suture clips. Mice were monitored for recovery, and suture clips were removed seven days postsurgery.
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