[Cytometry] Sorting KC167 Drosophilla cells

Rosemary Clarke r.z.clarke at dundee.ac.uk
Fri Jul 16 09:53:22 EDT 2010


Hi,

Has anyone out there any experience of sorting the embryonic  
Drosophilla cell line KC167? A sort that I thought would be easy is  
proving not to be, as the cells die on me. I have tried everything I  
can think of to improve things. Increased nozzle size from 70 micron  
to 90 micron (though cells are only 10micron in diameter). Sorting at  
RT and 4 degC. Sorting into FCS or 'normal' media. The intended sort  
was of cells loaded with either Dycycle Orange or Hoechst, and we  
thought it may be the dye but even unstained cells die. The only  
thing I can think that's left to change is the sheath fluid, which at  
the moment is PBS. Oh, and I am using a Vantage at 10 psi.

Any ideas out there?

Thanks in advance
Rosie

Dr Rosie Clarke
Division of Cell Biology and Immunology
WTB/MSI
University of Dundee
Dundee
Scotland
UK
Tel: (01382) 384222
E-mail: r.z.clarke at dundee.ac.uk




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