Re: [GFP Retention]

From: Jan Grawé (jan.grawe@rudbeck.uu.se)
Date: Fri Dec 21 2001 - 02:49:50 EST


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



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