Re: Effect of storage on retic staining
Bruce H. Davis, M.D. (bdavis@beaumont.edu)
Mon, 29 Jul 1996 17:28:39 -0700
Raymond B. Hester wrote:
> doing 2-color FACS (we have a 440) with thiazole orange (TO) to
> detect retics and a phycoerythrin conjugated antibody to look at another marker. The problem we are having is that
> sometimes the TO-only stained cells sometimes appear in the lower
right (530/30) quadrant as expected but sometimes they appear in the
two-color quadrant as if they were doubly stained.
Ray: The problem you describe is actually predictable based upon the
spectral properties of thiazole orange (see Lee et al, Cytometry 7:508,
1986 or editions of Howard Shapiro's excellent text). TO emits further
into the spectral range of PE relative to FITC. If you need to do two
color work with TO, you will need to use a Moab signal in the red range,
such as tri-color or PerCP.
Bruce H. Davis, M.D.
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