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>Corn pollen (from PolySciences), about 90um in diameter, was used as
>a large test particle. The roughly 60us pulse width for pollen was
>as expected (beam height is assumed to be 20um at focus). However,
>there were substantial problems with clogging, hence, a sample tubing
>with larger internal diameter (and about the same outside diameter)
>is needed. Any reliable sources?
BD makes larger sample tubing:
250 um i.d. 80-30059-00
500 um i.d. 80-30061-00
>Further, sample settling is, of course, a problem and the informal
>documentation recommends against using sample agitate while sorting,
>but suggests the addition xanthan gum at 0.1% concentration. Any
>experience with this vis a vis, fluidics effects (viscosity of the
>sample stream will go up), sorting efficacy, sterility of sorted
>cells?
I have heard the same from BD, and they swear by it (!), although I haven't
tried it yet.
>Any other gems of wisdom in sorting large particles on the FACStar?
Just what I've been told (several times) from BD: if something doesn't look
right, there are probably air bubbles in the flow tip. Use the syringe trick to
get rid of bubbles FIRST, before trying anything else.
The "Sort Setup" and "MacroSORT User's Notes" from training provide
'cookbook'-style instructions and hints that I find useful.
We haven't used the 'hoses' to sort basketballs yet, so all of this is 2nd-hand
information. :-) We routinely use MacroSORT with a 70um tip, however.
/\/\/\_ Eric Van Buren vanburen%flovax.dnet@rocdec.roc.wayne.edu
\ \ \ Immunology & Microbiology
\_^_/ Wayne State University
...first there was Cytometry 2000, then GLIFCA, and now...GLIFCA 2 (with SCFCA)!
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