Mouse Splenocyte and splenic lymphocyte count - (May/09/2007 )
Dear all,
I 've been preparing mouse splenic lymphocyte from 6-8 weeks female BALB/c mice using nylon mesh and ficoll paque centrifugation and my cell count is only 1.3-1.8x10e7 per spleen (trypan blue). But when I count before ficoll, its around 8x10e7. Does this figure sound right? What critical step should I pay special attention to improve this?
Many thanks!
Kallichroma
-Kallichroma-
QUOTE (Kallichroma @ May 9 2007, 07:36 AM)
Dear all,
I 've been preparing mouse splenic lymphocyte from 6-8 weeks female BALB/c mice using nylon mesh and ficoll paque centrifugation and my cell count is only 1.3-1.8x10e7 per spleen (trypan blue). But when I count before ficoll, its around 8x10e7. Does this figure sound right? What critical step should I pay special attention to improve this?
Many thanks!
Kallichroma
I 've been preparing mouse splenic lymphocyte from 6-8 weeks female BALB/c mice using nylon mesh and ficoll paque centrifugation and my cell count is only 1.3-1.8x10e7 per spleen (trypan blue). But when I count before ficoll, its around 8x10e7. Does this figure sound right? What critical step should I pay special attention to improve this?
Many thanks!
Kallichroma
I don't do ficoll paque as you will lose lots of lymphocytes.
You can get rid of red blood cells by lyzing them with NH4Cl for 5min in 37degree water bath.
Hope this may help.
-Minnie Mouse-
We also do not do Fiq. Hq but after making a cell suspension, perfom ACK lysis to get splenocytes.
-Nabin-