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> Does anyone have any experience using HBSS (Hanks Buffered Salt Solution)
> as sheath in a sorter?
[snip]
> it clogs our sheath filter.
We use home-made isotonic Na/K saline with Tris buffer, Ca++ and Mg++. We
have limited sorting experience but we've done lots of analysis (FACStar
and FACScan) without any filter clogging; we're not aware that this has
caused any sorting problems. HBSS per se will climb to a rather high pH in
air due to loss of carbon dioxide. At this pH, there may be
microprecipitation of e.g. calcium hydroxide. Keeping the pH reasonable
with HEPES would be rather $pendy; Tris would be cheaper but may be more
toxic. That's why we make up our own and avoid the bicarb altogether.
Also the HBSS pH instability could cause the FITC emission intensity to
drift. We did have problems with slow precipitation of phosphate till we
substituted Tris.
We add azide only when viable cell recovery is not an issue, and for
nonsterile storage of 10-liter stock carboys.
/*- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Eric Martz, Professor of Immunology emartz@microbio.umass.edu
Dept Microbiology Voice: 413-545-2325 FAX: 413-545-1578
Morrill IVN 203, Box 35720, Univ Massachusetts, Amherst MA 01003-5720
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