How to anesthetize mice ? - for both peritoneal and brain macrophge harvest (Dec/01/2007 )
Hi guys
i have a quite argent problem regarding mice anesthetization. i doing an In-vivo experiment and i need to anesthetize the mice for both harvesting of micrgolia and peritoneal macrophages.
when i harvest microglia i anesthetize the mice using ketamin (i.p injection) and when i'm harvesting peritoneal macrophages i'm using CO2. but what do when i need both of them (i fear that an i.p. injection will harm the peritoneal macrophages and that CO2 will cause harm to the microglia....)
anu ideas ??? please help !!!!
L.
What about isoflurane?
yes, many labs use gaseous anesthetics like isoflurane during rodent surgery.
I think that for isoflurane you have to use it in a mixture with N2. and in any case we are out of Iso and the experiment is on Monday (and i can't postpone it).
I found this paper "Normal Adult Ramified Microglia Separated from Other Central Nervous System Macrophages by Flow
Cytometric Sorting" form The lournal of Immunology, 1995 in which they used CO2 asphyxiation and than harvested microglia - so i'm hoping using Co2 will be OK. what do you think ? have any experience with it ???
are you intending to sacrifice the mice... or keep them alive after?
because if you intend to sacrifice them... cervical dislocation would be the best bet.
short term sleepy time: co2 should be ok. never used it though.
http://www.lal.org.uk/pdffiles/Co2.pdf
V
because if you intend to sacrifice them... cervical dislocation would be the best bet.
short term sleepy time: co2 should be ok. never used it though.
http://www.lal.org.uk/pdffiles/Co2.pdf
V
they will be sacrificed.... and i'm afraid that cervical dislocation will cause bleeding (in the CNS)
Ok....
i couldn't use Co2 with some experiments because it "potentially" interferred with some gene expression, hence cervical dislocation.
we do use CO2 for permanent sleepy time, when it's appropriate. Place the mice in a cage, place a lid attached to the CO2 hose, and turn the tap on. I do not recommend looking at the mice as they breathe the gas in. Check for responseby pinching their toes.
V
I've crossed an article which states Isoflurane treated cells develop AD or Amyloid ß level is elevated..