Another means of studying the nature of the granules depends on the
principle of =vital staining=. The "vital methylene blue staining"
(Ehrlich) that has since become so important, especially in neurology,
led to the first attempts at staining the granules in living animals.
One of the first publications on this subject is that of O. Schultze,
who placed the larvae of frogs in dilute methylene blue solution, and
after a short period found the granules of the stomach, the red blood
corpuscles and other cells stained blue. This method, however, cannot
pass as entirely free from error, as Ehrlich frequently found that when
the experiment lasts some time the methylene blue often forms granular
precipitates that may be confused with the granules. Teichmann directs a
detailed analysis to this point, and regards most of the granules
described by Schultze as artificial products.
=Neutral red= is highly suitable for the study of vital granule-staining,
a dye recommended by Ehrlich, and employed successfully since that time
by Przesmycki, Prowazek, S. Mayer, Solger, Friedmann, Pappenheim and
others. This dye was prepared by O. N. Witt from nitrosodimethylamin and
metatoluylendiamin, and is the hydrochloric acid salt of a base which is
soluble in pure water, yielding a fuchsin red colour, but which in weak
alkaline solution--the alkalinity of mineral water suffices--is a
yellow-orange hue.
Now neutral red is characterised by a really maximal affinity for the
majority of the granules. Ehrlich was able by the aid of this dye to
demonstrate granules, even in some vegetable cells. Moreover the method
of using it is the simplest conceivable, as subcutaneous or intravenous
injection, or even feeding, in the higher animals stains the granules;
with frog's larvae and invertebrates, to allow them to swim in a dilute
solution of the dye is often sufficient. The staining also succeeds in
"surviving" organs, and is best effected by allowing small pieces to
float in physiological salt solution, to which a trace of neutral red is
added, under plentiful access of air. When the object is macroscopically
red it is ready for examination.
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