[Cytometry] Sorting KC167 Drosophilla cells]

Rochelle Diamond diamond at caltech.edu
Sun Jul 18 13:03:39 EDT 2010


Dear Rosemary,
For primary drosophilia embryo cells we use Seecoff saline as the sheath
fluid.  The recipe is as follows:
6mM Na2HPO4
3.67 mM KH2PO4
106mM NaCl
26.8mM KCl
6.4mM MgCl2
2.25mM CaCl2
pH6.8

We get much improved viability with this.

Best regards,
Rochelle (Shelley) Diamond
Caltech Flow Cytometry Cell Sorting Facility
Pasadena CA USA

On Fri, July 16, 2010 6:53 am, Rosemary Clarke wrote:
> 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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