FREEHAND SECTIONS 
89 
1 or 2 minutes in 95 per cent alcohol to remove the acid. A trace of 
sodium carbonate, just enough to make the alcohol alkaline, may 
be added to the 95 per cent alcohol. If any acid remains, the 
safranin will fade. Dehydrate in absolute alcohol 1 to 5 minutes, 
clear in xylol, or first in clove oil and then in xylol, and mount in 
balsam. 
For convenient reference, the process may be summarized, but 
it must be remembered that all the schedules are intended merely to 
introduce the method to the beginner. 
1. Sections in 95 per cent alcohol. 
2. Stain in safranin, 24 hours. 
3. 50 per cent alcohol until the stain becomes weak in cellulose walls, but 
not until it is removed entirely. 
4. Anilin blue, 2 to 10 minutes. 
5. 95 per cent alcohol, 2 to 5 seconds. 
6. 95 per cent alcohol, slightly acidulated with hydrochloric acid, 5 seconds. 
7. 95 per cent alcohol, with or without a trace of sodium carbonate, 1 or 2 
minutes. 
8. Absolute alcohol, 1 to 5 minutes. 
9. Xylol, 1 to 5 minutes. The xylol may be preceded by clove oil. 
10. Mount in balsam. 
Lignified and suberized walls should stain bright red and cellulose 
walls bright blue. To make this beautiful combination a success, it 
is necessary to be very careful. If too much safranin is extracted 
at stage 3, the acid at stage 6 will still further weaken the red stain 
and the contrast will not be sharp. 
Safranin and Light Green (Land’s Schedule).—This is another 
beautiful combination and the student should be successful from 
the first, since the light green is simpler to apply than either Dela- 
field’s haematoxylin or anilin blue. 
Land uses either aqueous, anilin, or alcoholic safranin, and 
uses the light green in clove oil, or in a mixture of clove oil and 
absolute alcohol. Make a saturated solution of light green in clove 
oil. Since the solution takes place slowly, the mixture should stand 
several days before using. If a small quantity of absolute alcohol 
be added to the clove oil, the stain dissolves more readily. For 
some structures the stain is more brilliant than with the simple 
clove-oil solution. 
