>Hi, Everyone,
>I am doing a screening with fluorescence-activated cell sorting 
>technique. Even after optimization (plus sucrose in the collection 
>tube suggested by Adam Kuspa, rescue and grow cells on KA plate), 
>only ~10% of cells survive through this physical sorting. This makes 
>large-scale screening more difficult.
>The cells were sporulated for 24 hours, and then applied to FACS machine.
>Any suggestions for further cell viability are welcome.
>Thanks, Xin-Hua Liao


Dear Xin-Hua Liao,

Dictyostelium cells turn out to be very sensitive to pressure in the 
feeder pipes of FACS machines. At a pressure used for lymphocytes (70 
psi, pounds per square inch) most vegetative Dictyostelium cells die. 
We now routinely use 9 psi, with good subsequent viability.

Good luck for your experiments,

Pierre



-- 
Pierre Golstein
Centre d'Immunologie INSERM/CNRS/Univ.Medit.
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