Fix in 1 % paraformaldehyde before sorting?
Jan Grawé wrote:
>
> James,
>
> In order to reduce salt buildup on slides when sorting rare events I
> have on a FACStar+ routinely sorted using PBS diluted 1:300 to 1:500.
> 1:1000 dilution was (on our instrument) also possible , but somewhat
> unreliable.
>
> You would propably have to adjust the droplet charge, pulse shape,
> and phase significantly in comparision to sorting with PBS.
>
> Jan Grawé
>
> >Hello,
> >
> >We have a client who is trying to sort a fresh water protozoan that
> >expresses GFP. They are very worried about stressing the cells. They
> >have found that stressing these cells causes them to release their
> >GFP and thus show up negative. I was wondering if anybody has
> >determined the minimum concentration of salt that can be used in the
> >sheath fluid that will still hold a charge. Presently we use a 1X
> >PBS solution consisting of NaCl, KCl, Na2HPO4 7H2O, and KH2PO4
> >
> >
> > Thanx in advance, Marv
>
> --
> --------------------------------
> Jan Grawé
> Cellanalyslab
> Rudbecklaboratoriet/C5
> Uppsala Universitet
> Dag Hammarskjölds väg 20
> 75185 Uppsala
>
> tel: +(0)18-4714656
> mobil:+(0)70-2577874
> fax: +(0)18-552739
> epost: jan.grawe@rudbeck.uu.se
> http://www.rudbeck.uu.se/cellanalys
--
________________________________________________________
/ Michael J. Herron, U of MN, Dept. of Pediatrics/BMT /
/ herro001@umn.edu /
/ 612-626-4321 Mpls MN 55455 /
/_______________________________________________________/
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