Optimal sheath -condishioned media??

jim_phillips@mednet.med.miami.edu jim_phillips at mednet.med.miami.edu
Fri Aug 7 16:10:41 EST 1998



     
        Hello to all, 
        This is just a quick note about cells not liking 
        being put in a 96 well by themselves.  I suggest 
        using what we term as "conditioned media".  It is 
        essentially used media.  Take some media from a 
        passage (spin it hard) and either add (or not) a  
        percentage of new media to the 96 well plate to make 
        the cell think that it is not alone.  This may or 
        may not help in all cases but it is worth a try if 
        your cells keep dying.
        
        Jim Phillips
        University of Miami School of Medicine

______________________________ Reply Separator _________________________________
Subject: Re: Optimal sheath fluid for cell sorting 
Author:  "Steve G. Hilliard" <steve at habanero.cb.uga.edu> at SMTPMED
Date:    8/7/98 9:11 AM


     
Simon,
     
This was one of our concerns, but our calculations indicated that 
each drop would contain a fraction of a microliter (1/20th?), so 
if you're doing single cell sorts for cloning the volume of PBS you're 
adding to your media should be insignificant.  If you're sorting higher 
numbers then it might add up, but with a cushion of 100-200ul of media 
you're still talking a 1:100 ratio.  
     
You could sort into media w/ higher than normal serum content if you're 
really concerned, but I would not expect problems.  The main problem is 
that cells don't grow well when they're lonely! ;-(
     
Good luck,
Steve
     
-------------------------------------------------------------
    Steve G. Hilliard                  flowman at uga.edu        
       University of Georgia Cell Analysis Facility 
             http://floweb.cb.uga.edu/Floweb/
-------------------------------------------------------------
     
     
On Thu, 6 Aug 1998 Simon_Q_Rice at sbphrd.com wrote:
     
> Dear Flowers,
> A question for the cell sorters amongst you..... 
> 
> I am sorting cultured cells into 96-well plates (wells containing
> appropriate culture medium) and am currently using PBS as a sheath fluid. 
> Experience tells me that cultured cells aren't too keen on PBS for long
> periods of time and I wondered if this could have a deleterious effect on 
> the cells.  Therefore, what alternative sheath fluid, if any, would you
> recommend?
> 
> Apologies if this is a naive question. 
> 
> Thank you in anticipation
> 
> Simon
> 
> 
> 
> 
> 
> -- End --
> 
     
     





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