20 
Giemsa's Stain 
1 part), and may be left in this for twelve hours with advantage. If 
preferred the fixation may be completed in aqueous Sublimate solution, 
but then the slide must be treated for some time with 70 Vo Alcohol, as 
otherwise the corpuscles are liable to become detached from the glass 
during staining. 
The remaining processes are the same as those given by Giemsa 
{Deutsche med. Wochenschr. No. 40. 1909, and No. 12. 1910) for the 
application of his stain to sections. 
4. The slide is washed in several changes of distilled water and then 
placed for ten minutes in the following solution 
Potassium iodide 2 gms. 
Lugol’s solution 3 c.c. 
Distilled water 100 c.c. 
(Lugol’s solution consists of KI 6 gms.. Iodine 4 gms. distilled water 
100 c.c.) I consider that the ordinary Gram’s Iodine solution acts equally 
as well as this mixture, 
5. After washing for a short time in water, the slide is placed for 
ten minutes in a 0 5 "/o solution of Sodium thiosulphate, then washed 
for five minutes in tap water, and for five minutes in several changes of 
distilled water. 
6. It is then stained for from two to twelve hours in a solution of 
one drop of Giemsa’s solution (Griibler) to 1 c.c. of distilled water. It 
is essential that this distilled water be as neutral and as pure as 
possible. As a rule ordinary distilled water cannot be used as it often 
contains organic acids and other impurities. Such water should be 
treated with Potassium bichromate and Sulphuric acid, and re-distilled 
in an “all glass” still and condenser. 
A fresh solution of the stain may with advantage be used after the 
first half hour. 
7. After staining, the slide is rinsed in re-distilled water, and 
differentiated in the following solutions 
1. Acetone 95 parts, Xylol 5 parts. 
2. Acetone 70 „ Xylol 30 „ 
3. Acetone 70 „ Xylol 30 „ 
4. Xylol, pure. 
It is necessary that the Acetone and Xylol be pure and absolutely 
neutral. Finally the slide may be examined in Cedar Oil, or mounted 
in neutral Balsam in Xylol. This neutral Balsam solution can be 
