Is there a way to induce hypoxia without a chamber, for example using a chemical such as sodium azide?
Scott et al. Science (2002) describe the use of 0.5M NaAz for 1.5hrs. However following this protocol (and in attempting to optimize it) I have been unable to obtain viable animals.
Does anyone know of a different method for inducing hypoxia?
Thanks and kind regards,
Brent
You could try cobalt chloride (CoCl2). Cobalt chloride mimics hypoxia in mammalian cells, and I think also in S. pombe.
http://scholar.google.com/scholar?q=cobalt+chloride+hypoxia&hl=en&lr=&btnG=Search
-Cory
We use 0.5M azide for one-hour and get anywhere between 20-80% killing at room temperature. It’s really important to be consistent and to wash the worms several times after incubating them to get rid of the azide. I think that there’s alot of variability inherent in the protocol…look at the Dasgupta paper from Crowder’s lab on HPC…their paired trials are all over the board, though they see the same effect or trend in each pair (ie +/- HPC). 1.5 hrs is too long, though, at least in our lab. Good luck.
KWN