Re: Characteristics of eosinophils and basophils
Howard Shapiro (hms@shapirolab.com)
Mon, 3 Jun 1996 10:57:40 -0400 (EDT)
Mike Loken, Leon Terstappen, et al showed antigenic characteristics of eos
and basos in their older (mid-to-late-'80's) work on doing flow differential
counts, with antibodies plus the nucleic acid dyes thiazole orange and LDS 751.
Light scattering characteristics of these cells can vary somewhat with
preparation; unfixed, unstained eosinophils have orthogonal scatter signals
toward the high end of the granulocytes but do not form a distinct cluster,
whereas glutaraldehyde fixation moves the eosinophil cluster away from the
neutrophils. Basophils tend to have low orthogonal scatter values, and to
fall among the lymphocytes on a forward vs. orthogonal scatter plot, but
scatter signals may be affected by degranulation, which occurs in the
presence of at least some lysing agents, such as NH4Cl. The best way to
identify unfixed, unstained eosinophils is probably on a plot of polarized
vs. depolarized orthogonal scatter as described by Terstappen, De Grooth, et
al.; I don't know to what extent fixation alters the birefringence of
eosinophil granules which accounts for the high depolarized scatter signals.
-Howard
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