Hi everybody,
Can anyone describe the exact protocol to set up this animal model of retinal degeneration?
My questions are as simple as:
(i) When you put back to normoxia the mice at P12 (after the 5-days period of 75% hyperoxia), do you just take the cage outside the oxygen chamber back to the atmosphere normal room conditions or do you keep them in the cage and set up the oxygen rate to normoxia?".
(ii) Could you provide me a list of the material I need to set up this model?
(iii) Are there any trick I should know before starting?
(iv) Are C57BL/6 sensitive to this treatment?
(v) How do you manage the death of the feeder mother? I mean do you specially buy some feeder mice for each experiment or do you use some feeder mice from you animal facility?
Thanks a lot for all the answers to those questions and for all the détails you could bring into my knowledge!
Juliette
I'm actually looking into establishing the same model. So a littlbe bit unsure what materials shuld be used with reasonable cost\perfomance results. I have some knowlege about this model as other people in my old lab was working with it.
(i) When you put back to normoxia the mice at P12 (after the 5-days period of 75% hyperoxia), do you just take the cage outside the oxygen chamber back to the atmosphere normal room conditions or do you keep them in the cage and set up the oxygen rate to normoxia?".
It's pretty much depends on your equipment if you can to change the gas composition in the cage to normoxia and do it slowly that you definitly shuld do it before returning the cage to normal enviroment. If you can't than you simply stop the oxygen flow and unseal the cage or container letting the normal atmosphere to slowly diffuse into the cage.
(ii) Could you provide me a list of the material I need to set up this model?
1. Airtight chamber with cage inside or special cage http://www.coulbourn.com/category_s/260.htm this company provide products for metabolical study in mice and rats but also can be used for OIR however I still not sure about the cost of this equoment.
2. 100% oxygen with gas mixer (if you don't have it try to get 75% oxygen\nitrogen mix then you can just perfuse the cage with this mix.
3. Gas analyser to make shure that enviroment in the cage is apropriate for the model.
(iv) Are C57BL/6 sensitive to this treatment?
Yes they are.
(v) How do you manage the death of the feeder mother? I mean do you specially buy some feeder mice for each experiment or do you use some feeder mice from you animal facility?
No, you get the mouse pregnant let the pups to be born in the cage, then you start the oxygen flow. You can't use any other mouse to feed your pups it will kill it instead. It might also happen if you touch the pups, so the better way for a mouse to give a birth inside the cage that later would be subjected to oxygen flow.
Please let me know if you found any practical solution for oxygen perfusion and the estimate cost of it.
I have found that it is potentially possible to use airtight cages for mice and perfuse them with oxygen under went in order to not fill the lab room with oxygen. I'm still looking for a cheap and good oxygen analyser for this work.
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