isoflurane and FACS analysis of peripheral blood?
Posted 16 May 2009 - 09:10 PM
i have to use an inhalant anaesthetic because my collaborator is taking the brain for electrophysiological recordings.
we anaesthetise the mouse in 2% isoflurane for a few seconds, and then we perform decapitation. I obtain the peripheral blood in EDTA (0.5%) pretreated blood, perform a RBC lysis (BD Pharmalyse) for 5 minutes, and wash the cells with PBS at 1200rpm for 6 minutes.
when i try the protocol on mice without isoflurane (but CO2 for example) the blood looks perfect in FACS - all the cell populations are seen.
is there anything you recommend? does anyone know how isoflurane is effecting blood?
Posted 18 May 2009 - 11:08 AM
Posted 18 May 2009 - 12:00 PM
Vet Res Commun. 2008 Dec;32(8):619-26
J Appl Genet. 2005;46(3):319-24
Int J Immunopharmacol. 1997 Nov-Dec;19(11-12):699-707
Another option for anesthesia could be intraperotineal avertin. There's precedent for using cells from rats sacrificed this way for EP. Check out:
Nature Protocols 2007 Vol. 2 No. 1 pp. 152-160
then heaven will be yours, before you meet your end
Posted 01 June 2009 - 02:35 AM
One thing I found helpful is diluting the blood sample 1:1 with PBS to increase fluidity of sample.
Posted 11 June 2009 - 10:40 AM
Posted 17 June 2009 - 06:50 AM