[Histonet] question about mouse perfusion
dancingwing <@t> yahoo.com
Wed Mar 17 13:58:00 CDT 2010
I have just started to learn how to do perfusion in mice and this site has helped me a lot.
We don't have the pump or Perfusion one in the lab, and probably won't be able to get one in the near future, so I'm using the hand injection method. I'm using a 60 ml syringe with 21G needle. I am doing perfusion in order to do immunofluorescence on the brain.
My question is: can anyone give me a rough idea of how fast I should inject ( like ml/min). I think I've tried like 30 ml in 3 min, and I suspect that it's too fast because I do observe tissue swelling sometimes.
And another question, which of the following is better for perfsusion: 1) PBS and then 4% PFA 2) Saline and then 4% PFA 3) sucrose--> saline--> PFA?
Thank you very much for your help!
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