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Basophil counting with a new staining method using alcian blue
HS Gilbert and L Ornstein
Difficulties in obtaining reproducible and accurate enumeration of
circulating basophils with existing techniques have hampered investigation
of this infrequent cell population. A new basophil staining method is
described that employs alcian blue dye for staining of heparin within
basophils at low pH and in the presence of lanthanum ions. Basophil
recognition is facilitated by reducing nonspecific nuclear staining. This
objective is achieved because of the differences in stability of alcian
blue-heparin, alcian blue-nucleic acid, lanthanum-heparin, and
lanthanum-mucliec acid complexes. Reduction of pH after staining also
favors solubilization of leukocyte cytoplasmic proteins, providing greater
contrast between stained and unstained cells by reducing light scattering
of the unstained leukocytes . The alcian blue staining method is suitable
both for chamber basophil counting and automated basophil counting using
continuous-flow sampling and electro-optical detection. The new staining
method was evaluated by comparing it with the chamber counting method using
toluidine blue in a triple-blind study in which the results of basophil
counting by the alcian blue chamber method, alcian blue automated
instrument method, and the toluidine blue chamber method were analyzed for
reproducibility and compared with an indirect basophil count obtained from
a 1000-cell leukocyte differential and a total leukocyte count. Both alcian
blue staining methods gave greater reporducibility that toluidine blue and
were more accurate, as evidinced by a significantly higher correlation with
the indirect basophil count. The improved reproducibility, accuracy, and
convenience of this method over existing methods should facilitate the
collection of more meaningful information about circulating basophil levels
in health and disease.
Volume 46,
Issue 2,
pp. 279-286,
08/01/1975
Copyright © 1975 by The American Society of Hematology
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