To 1 cc. of distilled water in a small, perfectly clean graduate
add 1 drop of the stain, shaking very gently. Make very thin
film; dry in air; fix 15-20 minutes in absolute alcohol. Cover
preparation with a thin layer of the freshly diluted stain for
10-15 minutes, renewing stain at end of 10 minutes. Wash in a
stream of water. Differentiate over-stained preparations in
distilled water. Dry with absorbent paper; mount in balsam. Stains
the _spirochæte pallida_ and _malarial organisms_. The Giemsa
solution may be obtained from Grübler. A more intense staining can
be obtained by adding to the water used for diluting the stain 1-2
drops of a 0.1 per cent solution of potassium carbonate.
5. SPECIAL ELECTIVE STAINS FOR THE BLOOD.
1. =Ehrlich’s Triple Stain.=
Saturated watery solution of Orange G 120 cc.
Saturated watery solution of acid fuchsin 80 cc.
Saturated watery solution of methyl green 100 cc.
Glycerin 50 cc.
Distilled water 300 cc.
Absolute alcohol 180 cc.
Mix gradually; allow to stand for several months; do not shake or
filter. Remove stain with pipette. Fix by heat, or pure methyl
alcohol for 5 minutes. Stain 5-10 minutes; wash thoroughly, dry
and mount in balsam. Neutrophile granules violet; eosinophile,
a bright red; nuclei of the neutrophilic and eosinophilic cells
greenish-blue; nuclei of the lymphocytes deep-blue; nuclei of the
large mononuclears pale blue; those of red cells intense blue; red
cells copper red. The Aronson-Philipp modification is more variable
and less satisfactory.
Pappenheim’s Stain for Lymphocytes.
3-4 parts of polychrome methylene-blue or methyl green to 1-2 parts
of pyronin. Fix in absolute alcohol. Nuclei blue-green; protoplasm
bright red.
Staining of Blood-platelets.
The blood-platelets may be examined in the fresh state by coating
a cover-slip with Deetjen’s agar-solution (boil 5 grms. agar-agar
in 500 cc. distilled water, filter hot, and to each 100 cc. of the
filtrate add 0.6 grm. sodium chloride, 6-8 cc. of a 10 per cent
solution of sodium phosphate and 5 cc. of a 10 per cent solution
of sodium diphosphate). Place drop of blood on this coating and
examine on warm stage. For permanent stained preparations bleed
into a fixing and staining fluid (equal parts alcohol and ether and
Romanowsky’s stain) or use Wright’s stain.
Bremer’s Diabetic Reaction.
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