RE:Sheath fluid

From: Arnold Pizzey (a.pizzey@UCL.ac.uk)
Date: Wed Aug 29 2001 - 10:05:51 EST


Greetings Ole,

As far as I am aware, Hanks Buffered Saline Solution should be pH 7.4
If the stuff you are using isn't, try this recipe:
0.40 g KCl (5.4 mM)
0.09 g Na2HPO4x7H2O (0.3 mM)
0.06 g KH2PO4 (0.4 mM)
0.35 g NaHCO3 (4.2 mM)
0.14 g CaCl2 (1.3 mM)
0.10 g MgCl2x6H2O (0.5 mM)
0.10 g MgSO4x7H2O (0.6 mM)
8.0 g NaCl (137 mM)
(1.0 g D-glucose (5.6 mM))

I would be a bit wary of including the Glucose myself; if your cells
absolutely need glucose I would add it into the receiving tubes -or add
glucose sequentialy through the sort.

Adjust pH to 7.4 and add H2O to 1 liter and
Filter sterilize

Store indefinitely at 4°C

If you find HBSS better for viability, let us know.

Regards,


Arnold






At 03:10 PM 8/24/01 -0400, you wrote:
>         Sheath fluid for cell sorting Recently I was told that HBSS is
>preferable to regular PBS based sheath fluids. It is supposed to increase
>viability in sorted cells. Our problem is that HBSS is buffered with
>Na-Bicarbonate, it also contains glucose. If we made up a 10-liter batch
>and let it sit, it would turn alkaline. With the glucose as a nutrient
>there is also an increased risk of growth. We would like to know what the
>general opinion is on this.     Ole J. Bergh, Supervisor  Flow Cytometry
>Facility  University of Pittsburgh Cancer Institute  200 Lothrop Street W
>1009 Biomedical Science Tower  Pittsburgh, PA 15213  Phone:412 624 0399
>Fax: 412 624 9624  mailto:berghoj@msx.upmc.edu
>http://pci.upmc.edu/internet/flowcyto/flow.htm




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	Arnold Richard Pizzey
	Department of Haematology
	Royal Free and University College London Medical School
	98 Chenies Mews
	London WC1E 6HX
	U.K

	voice:	+44 020-7679-6234
	Fax:	+44 020-7679-6222
	email:	a.pizzey@ucl.ac.uk
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