Re[2]: Sorting Sheath Fluid- The easy Method
jphillip@mednet.med.miami.edu
Mon, 15 Apr 96 09:23:15 EST
I have found that the easy method to sorting sheath fluid is to
require every investigator that wants to sort to bring their own
sheath fluid. I had an experience about 9 years ago when I used to
provide the sheath fluid where someone claimed that my sheath fluid
was hypertonic and killed their cells. I solved this problem from
that day onward by making everyone bring the sheath fluid where their
cells are happiest. Everyone that wants to sort brings 2 liters of
their own sterile sheath, but no fetal calf in the solution because it
gets stringy and clogs the nozzle. I have been doing this for 9 years
and no one seems to mind, and you don't have to worry about osmolarity
and the people are happy because their cells are in the best solution
possible for their cell line or whatever. As I mentioned, this is the
easy method.
Jim Phillips
University of Miami (Suntan U.)
______________________________ Reply Separator _________________________________
Subject: Re: Sorting Sheath Fluid
Author: dick stovel <STOVEL@Beadle.Stanford.EDU> at smtpmed
Date: 4/13/96 6:19 PM
Lorie Miller asks:
> We normally sort using Haema-Line 2 as our sheath fluid in our
> FACStarPLUS. One of my users needs to sort but cannot have potassium
> or phosphate in the system. Any suggestions?
>
> What concentrations of NaCl or anything else are necessary for proper
> charging of the drops for sorting?
We used normal saline (.9% - same stuff they plugged all the hospital
patients into) happily for many years before switching to PBS
(Phosphate Buffered Saline) for our sheath fluid.
As for concentration of NaCl for electrical conductivity for sorting - not
to worry. Water with 1/50th of the concentration of normal saline will
happily conduct your drop charges.
Richard Stovel stovel@darwin.stanford.edu
Herzenberg Laboratory and Stanford Shared FACS Facility
Stanford University, Stanford CA
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