[Histonet] question about mouse perfusion

[Li Zhang]

Dear all,

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!