Cells were cultured in Dulbecco’s modified Eagle’s medium (low glucose, Wako Pure Chemical Industries) supplemented with 10% fetal bovine serum. Cells were maintained at 37°C in a humidified incubator supplied with 5% CO2. Irradiation with γ-rays was performed using a Cs-137 Gammacell 40 Exactor (NORDION, Canada) at a dose rate of 0.7–0.9 Gy/min, hydrogen peroxide (H2O2) (Wako Pure Chemical Industries) diluted with phosphate buffered saline (PBS), 1 M N-acetyl-L-cysteine (NAC) (Nacalai Tesque) diluted in distilled water or 100 mM caffeine (Wako Pure Chemical Industries) diluted in PBS under their respective conditions. Details of the treatment conditions are described in the results or figure legends.
Cs 137 gammacell 40 exactor
Manufactured by Nordion
Sourced in Canada
The Cs-137 Gammacell 40 Exactor is a self-shielded irradiator designed for reliable and consistent radiation exposure. It utilizes a Cs-137 radioactive source to generate gamma radiation. The Gammacell 40 Exactor is a compact, user-friendly device intended for research and industrial applications requiring controlled radiation exposure.
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3 protocols using cs 137 gammacell 40 exactor
1
Cell Culture and Stress Induction
2
Zebrafish Spermatocyte Chromosomal Spreads
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Adult zebrafish were placed in a plastic box, with water depth of 2 cm. Animals were exposed to γ-rays for 11 minutes 42 seconds at 0.855 Gy/min (total exposure 10 Gy) in a Cs-137 Gammacell 40 Exactor (MDS Nordion). Irradiated fish were anesthetized 30 minutes after irradiation, and spermatocyte chromosomal spreads were prepared as indicated above. For each genotype, non-irradiated siblings were processed as controls.
3
Quantifying DNA Damage in C. elegans
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For DAPI body staining, late L4 larval stage worms were exposed to γ-rays for 58 min 30 s at 0.855 Gy/min (total exposure 50 Gy) in a Cs-137 Gammacell 40 Exactor (MDS Nordion, Ottawa, Canada). Irradiated worms were fixed 18–22 hr after irradiation for DAPI staining, and imaged to score DAPI-stained bodies as above. For western blotting of DSB-1, approximately 24 hr post-L4 worms were irradiated with either 10 or 100 Gy of γ-rays, and animals were lysed 1 hr post-irradiation.
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