How would you test the effect of antioxidants on C.elegans lifespan? Is this done through feeding E.coli with antioxidants and then C.elegans with E.coli, or is there another method?
base on a article they do the test of nano-Pt, kind of SOD, and they do the experiment in several steps, first choose C. elegans strains and growth conditions, meanwhile prepare the nano-pt. then, do the lifespan assay. thired is Oxidative stress resistance. the last step is Fluorescence microscopy. in the mictoscopy they do several steps, include Measurement of lipofuscin accumulation, Detection of ROS, Measurement of SOD/catalase activit, Thermotolerance and Effects of dietary restriction
You can simply add the molecule of interest in the NGM or Liquid medium.
From this paper.
The worms were bleach-synchronized as follows: 2 mL of 6% NaOCl were mixed with 1 mL of 5 M NaOH per 7.5 mL of concentrated worm suspension, and shaken for 4–7 minutes until the carcasses dissolved as monitored by direct observation. The remaining eggs were then washed 3 times by pelleting at ~1150 g for 2 minutes at room temperature, followed by aspiration of the supernatant and resuspension in 50 mL of 0.1 M NaCl. A final pellet of eggs was obtained by centrifugation at ~1150 g for 2 minutes at room temperature, followed by aspiration of the supernatant. Eggs were then added to a 250 mL liquid culture, as described above. For experiments without RNAi treatment, bacteria were heat killed at 80°C for 60 minutes. The worms were cultured at 20°C and monitored until they reached adulthood (~72 h), at which time FUdR was added to a final concentration of 400 µM. Worm viability was scored every two days by taking ten 10 µL drops (initially ~20 worms per drop) of the culture and counting the living worms under a microscope. The average number of living worms was then calculated. S medium or deionized water containing 10 mM malate, succinate, or fumarate was added back every three days to compensate for metabolism of the compounds and evaporation, and S medium containing FUdR and bacteria was replaced every 6 days. At least two replicates of each experiment were performed.