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1 Department of Anatomy, Harvard Medical School, Boston, Mass.
In acid solution, tissues tend to acquire a net positive charge and bind acid
dye; binding of basic dye is enhanced in basic solution where tissues tend to possess a net negative charge. A measure of the acid or basic strength of a given tissue
structure is afforded by its ability to bind basic or acid dye over a range of pH. The capacity of blood cell components to bind light green and methylene blue
in the pH range 0.05 to 12.3, and their affinity for eosin after acetylation and
treatment with nitrous acid, procedures used to block or destroy primary and
secondary amines and guanidino groups, was studied. Observations were also
made of blood smears stained with the Serra test for arginine. Erythrocytes bound light green through pH 12.3 and methylene blue to pH
6.0. The ground substance of white blood cells and immature blood cells stained
with light green through about pH 9 consistently, and often to pH 11: methylene
blue was bound through approximately pH 4.0. Eosinophilic granules bound
light green through pH 12.0, and did not bind methylene blue. Basophilic granules bound methylene blue in the pH range 8.0 to 1.0 but were dissolved in more
acid and basic solutions: they bound no acid dye. Neutrophilic granules were
stained with methylene blue through pH 5.3. Nuclei of immature blood cells
and monocytes bound methylene blue to about pH 4.0: polymorphous nuclei
and nuclei of lymphocytes and normoblasts were stained very faintly to pH
0.05. Many nuclei bound light green to about pH 12.0. Nuclear and cytoplasmic acidophilia was reduced or destroyed by acetylation
or treatment of smears with nitrous acid. Nuclei and cytoplasm of blood cells
were stained with the Serra test. Eosinophilic granules were stained strongly. The cytochemical significance of certain experimental findings have been discussed.
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