isolation of granulocytes - isolation of granulocytes (Feb/20/2003 )

I'd like to isolate granulocytes from whole rat blood or rat spleen. Does anyone have any experience with this ?

I am only used in isolation of rat granulocytes from pleural exsudates, following a carrageenan-induced pleurisy. It's very simple because you can have very quickly high amounts of "pure" granulocytes but you must keep in mind that the cells are activated for an inflammatory response...
However, I know that it is not easy to obtain them from blood samples because granulocytes are only a few percentage of the total cell content of the rat blood, in opposition to human blood. I have never try to obtain them by this way. If you need a protocol to try to isolate rat granulocytes with the pleurisy method or a method improved to human blood granulocytes, tell me...
Good luck...

I'm interested how to isolate granulocytes for luminol-dependent chemiluminescence tests... didn't find anything on the web... especially interested in centrifuging on ficoll-verografin gradient. if somebody can help in this, please mail me pecatum@yandex.ru