The Brooklyn Medical Journal. Vol. II. No. 2. Aug., 1888Various
Science
The Brooklyn Medical Journal. Vol. II. No. 2. Aug., 1888
Various
Medicine -- Periodicals
Now remove an amount which will just cover end of toothpick, and deposit
it on a previously cleaned cover glass, which should not be over 1/100
inch thick, and thinner if possible; immediately cover with another
cover glass, allowing sputum to spread by capillarity or slight
pressure, and separate by sliding apart, and put aside to dry without
heat. I have found that specimens dried without heat (and consequent
coagulation of albumen) will show a much larger number of bacilli than
when heat is used. I believe this is due to the fact that the fuchsine
penetrates more thoroughly through the albumen when not coagulated, or
that when it is coagulated by heat it to a greater or less extent it
protects them from the action of the stain. While the covers are drying
we will pour out a sufficient quantity of the aniline water, which by
this time has filtered into one of the staining glasses, and add one or
two drops (not more) fuchsine solution. Now, placing one of the cover
glasses on our cover holder, sputum side down, we lower it into the
staining fluid and withdraw holder from the side, and repeat the
operation for the other cover glass. It is my habit to allow the covers
to remain in this solution for at least eight hours or over night. The
time may be reduced to ten or fifteen minutes by heating the red stain
to about 140 or 150 F., but the result is not so brilliant, nor is it
sure, as I have frequently failed to find the bacilli by the short
method, but have been able to demonstrate their presence by the long
one.
At the end of either of the above periods of time, the cover glass is
lifted out of the staining solution and, without washing, immersed in
our five per cent. solution of nitric acid and alcohol. It is this part
of the process, if any, which will give trouble, as the time of
immersion is governed by the thickness and general character of the
sputum. My custom is to hold the first cover immersed until the color
has just disappeared, or say fifteen seconds, and the second five
seconds longer; but a very little experience will remove any difficulty
from over-decolorizing.
From the decolorizing solution they are immediately immersed in water
and thoroughly washed, when they may be again floated in the
contra-stain, which is prepared by filling the other staining glass with
water to which a few drops (three or four) of our methyl blue has been
added. They should remain here for from five to eight minutes, when they
are again removed with the pincetts, and a few drops of alcohol poured
over them to wash off the surplus stain. Again wash in clean water, and
dry by gentle heat (which will now do no harm) over the alcohol lamp,
and place sputum side up on table.
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