GFP tagged bacterial Cell preparation for FACS analysis - (Jun/19/2008 )
I want to analyse Corynebacterium glutamicum cells, which has GFP tagged with some gene in the genome, through FACS. But I am not sure about the cell preparation for FACS. Can anyone tell me the exact protocols.
Should I just harvest the cells and wash it with PBS thrice and store in fridge (4 degree) till I perform FACS.
How long I can store cells and what should be the concentration of the cells. How much sample is required for FACS. At what step cell counting should be done. Just before FACS or just after washing step.
Mahi
-mahi-
QUOTE (mahi @ Jun 19 2008, 08:58 AM)
I want to analyse Corynebacterium glutamicum cells, which has GFP tagged with some gene in the genome, through FACS. But I am not sure about the cell preparation for FACS. Can anyone tell me the exact protocols.
Should I just harvest the cells and wash it with PBS thrice and store in fridge (4 degree) till I perform FACS.
How long I can store cells and what should be the concentration of the cells. How much sample is required for FACS. At what step cell counting should be done. Just before FACS or just after washing step.
Mahi
Should I just harvest the cells and wash it with PBS thrice and store in fridge (4 degree) till I perform FACS.
How long I can store cells and what should be the concentration of the cells. How much sample is required for FACS. At what step cell counting should be done. Just before FACS or just after washing step.
Mahi
See if you find some answers in these links for bacterial flow cytometry: It would be good if you then post those answers here.
http://search.vadlo.com/b/q?sn=158621799&a...metry&rel=0
..
-cellcounter-
QUOTE (cellcounter @ Jun 20 2008, 02:02 AM)
It would be good if you then post those answers here.
-Bungalow Boy-