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>We have just sorted bacteria for the first time and since we got no
>growth when the sorted cells were placed back into culture, I thought I
>would run this by the group to see if anyone had suggestions.
The problem could easily be the sheath fluid.
Coulter Isoton II has antibacterial agents in it.
I am not sure of the exact composition but there are at least EDTA and
fluoride. There also seems to be something that gives the fluid a
distinctive smell, but it is not mentioned on the label.
I don't know how the cells were collected & stored before plating, but if
they were exposed to any significant concentration of sheath fluid they may
have been adversely affected.
Try sorting with plain PBS as sheath.
Dean R. Hewish, Cell Biologist & Flow Cytometrist.
CSIRO Biomolecular Engineering, 343 Royal Parade, Parkville, 3052,
Victoria Australia.
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