The cells were seeded into either a 6-well plate or a 10 cm culture plate and then irradiated with either X-rays or protons the next day. The irradiation was performed using the same plates, dishes and media volumes for both X-ray and proton irradiations. For X-ray irradiation, the cell dishes were located under a 2 cm thick bolus with a source surface distance of 100 cm and a field size of 30 × 30 cm2, and the cells were irradiated with 6-MV photons with a dose rate of 3.96 Gy per min as previously described [29 (link)]. The photons were delivered using a Varian Clinac 6EX linear accelerator (Varian Medical Systems, Palo Alto, CA, USA). For proton beam irradiation, the cell dishes were placed at the mid-spread-out Bragg peak (SOBP), and the cells were irradiated with a 230 MeV proton beam at a dose rate of 2.14 Gy per min generated by a proton therapy system (Sumitomo Heavy Industries, Ltd., Niihama, Japan) at the Samsung Proton Therapy Center in Seoul, South Korea. The proton beam was spread out using a ridge filter to a 10 cm wide SOBP. The dosimetry profiles and verification method of the proton beam were as previously described [21 (link)].
Clinac 6ex
Manufactured by Agilent Technologies
Sourced in United States
The Clinac 6EX is a medical linear accelerator designed for radiation therapy treatments. It is used to generate high-energy x-rays or electrons that can be directed to a patient's tumor or other target areas. The Clinac 6EX provides precise and accurate radiation delivery to treat a variety of cancers and other medical conditions.
Automatically generated - may contain errors
Lab products found in correlation
Truebeam, by Agilent Technologies (1 mentions)
Giemsa solution, by Kanto Chemical (1 mentions)
T25 flask, by Thermo Fisher Scientific (1 mentions)
Azd1775, by AstraZeneca (1 mentions)
Eclipse planning system, by Agilent Technologies (1 mentions)
Clinac 2100c, by Agilent Technologies (1 mentions)
Pinnacle planning system, by Philips (1 mentions)
Alamarblue assay, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by BD (1 mentions)
Annexin 5, by BD (1 mentions)
18 protocols using clinac 6ex
1
Comparative Radiation Dose Delivery Methods
2
Radiation Dose Delivery Verification
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BC cells were irradiated with clinical radiation beams as previously reported [10 (link),14 (link)]. For X-ray irradiation, the cells were placed under a 2 cm thick solid water phantom and exposed to various doses of 6 MV photons. X-rays were delivered using a Varian Clinac 6EX linear accelerator (Varian Medical Systems, Palo Alto, CA, USA) with a dose rate of 3.96 Gy/min. The absolute X-ray dose calibration was done based on a TG-51 protocol with Gafchromic film to a 1% accuracy. For proton irradiation, the cells were positioned in a mid-spread-out Bragg peak (SOBP) of 11.2 cm width and irradiated with 230 MeV proton beams at a dose rate of 2.14 Gy/min. Proton beams were delivered by a proton therapy machine (Sumitomo Heavy Industries, Tokyo, Japan) using the wobbling technique [34 (link)]. The absolute proton beam dose was verified to 1% accuracy using the TRS-398 dosimetry protocol. The physical proton dose was used for all experiments, and there was no correction for an RBE of 1.1, as is the clinical practice.
3
Radiosensitization of Prostate Cancer Xenografts
4
Nanoparticle-Mediated Radiosensitization Assay
5
Ferroptosis Induction and Radiosensitization
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Cells were irradiated with X-rays using a linear accelerator (CLINAC 6EX, Varian Medical Systems, Palo Alto, CA) at doses of 2.5, 5.0, 7.5, and 10 Gy (dose rate, 2.19 Gy/min). The prescribed dose was defined to be at the isocenter. HeLa and NCI-H1975 cells in 60 or 90 mm plastic dishes were allowed to adhere to the dishes at 37°C in 5% CO2 for 6 h. Subsequently, they were treated with erastin alone (0.1, 1, 5, 10, 20, or 50 μM) or with erastin and ferrostatin-1 (1 μM) and incubated for 24 h.
6
Clonogenic Assay for Radiation Survival
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Cell survival was measured using clonogenic assay in triplicates as previously reported [19 (link)]. Briefly, cells were irradiated with graded doses of 0, 2, 4, 6, and 8 Gy of 6 MV X-ray (Clinac 6EX, Varian Medical Systems, Palo Alto, CA). Cell survival data were fitted to a linear-quadratic (LQ) model [20 (link)]. Clonogenic assay was repeated three to four times for each cell line.
7
Irradiation of Tumor-Bearing Mouse Legs
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Tumors implanted into mouse hind legs were irradiated as previously described [22 (link)]. Briefly, 6 MV photon beams were delivered at a dose rate of 3.96 Gy/min using a Varian Clinac 6EX linear accelerator (Varian Medical System, Palo Alto, CA, USA). Prior to irradiation, mice were fixed by anesthetizing with intraperitoneal injections of 30 mg/kg zolazepam/tiletamine and 10 mg/kg xylazine. Tumor-beading hind legs were placed inside the radiation field (30 cm × 7 cm) and covered by a 2-cm-thick water-equivalent bolus with a source-to-surface distance of 100 cm. The TG-51 protocol was used for calibration of the absolute dose.
8
Evaluation of EPID Performance on Linear Accelerators
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All tests were conducted on four linear accelerators at the Crown Princess Mary Cancer Centre (Westmead, Australia), namely: two Varian Clinac® 6EX (Varian Medical Systems, Palo Alto, CA, USA) with the aS500 EPID, one Varian Clinac® 21iX with the aS1000 EPID, and one Varian Truebeam® with the IDU EPID. The sensitive area for all three panels is 40 cm × 30 cm. The EPID aS500 has an array of 512 × 384 pixels and the EPID aS1000 and IDU has an array of 1024 × 768 pixels. Pixel size for the aS500 is 0.784 mm and for both the aS1000 and IDU panels is 0.392 mm, i.e., half the pixel size of the aS500. However, the images acquired on the Clinac® 21iX (in dosimetric mode) are of lower resolution with 512 × 384 pixels and a pixel size of 0.784 mm, the same as for the aS500.
9
Clonogenic Assay: X-Ray Radiation Protocol
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The cultured cells were counted by using a hemocytometer (Erma, Tokyo, Japan) and plated in T25 flasks (156367, Nunc, Waltham, MA, USA). The cells were allowed to adhere overnight prior to irradiation. After exposure to 6-MV-lianc X-rays (Clinac 6EX, Varian, Palo Alto, CA, USA), cells were incubated for 14 days at 37 °C in a humidified atmosphere of 95% air 5% CO2. Colonies were fixed with methanol and stained with 2% Giemsa solution (Kanto Chemical Co. Inc., Tokyo, USA). When calculating surviving fraction, the colonies located in the penumbra regions (−1.0 < x [cm] < 1.0 in Figure S1 ) were excluded. The surviving fraction is the ratio of plating efficiency of the irradiated group to that of the non-irradiated group. The PHITS calculation showed that the out-of-field dose relative to the in-field dose is 5.0% on average (Figure S1 ).
10
Radiotherapy Dosimetry Protocol for AZD1775
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AZD1775 was obtained from AstraZeneca (AstraZeneca, Waltham, MA, USA). For in vitro experiment, dimethylsulphoxide (DMSO) at 10 µM was used for vehicle and control. IR was performed using a Varian Clinac 6EX linear accelerator (Varian Medical Systems, Palo Alto, CA, USA). Cell monolayers were treated with various doses of 6 MV photons at a rate of 3.96 Gy/min. The distance between the IR source and the cell plates were kept at 100 cm while the field size was fixed at 30 × 30 cm. Cell plates were placed under a 2-cm-thick solid water phantom. The absolute photon dose was calibrated according to TG-51 and verified with Gafchromic film to 1% accuracy.
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