APPENDIX II 689 
reader is referred to “Stain Technology,” published by the 
Commission on Standardization of Biological Stains, Geneva, 
Di Ys 
METHODS FOR THE PREPARATION OF CANADA BALSAM Mounts 
No. 1. SAFRANIN AND Fast GREEN 
1. Safranin. By means of an eye-dropper put 10 to 15 drops 
of a strong alcoholic solution of safranin into a watch glass of 
distilled water. Allow the sections to lie in this mixture until 
deep red. 
_ 2. Transfer the sections to another watch glass containing tap 
water and rinse off excess of stain. 
3. Dehydrate and differentiate in 50 per cent. alcohol. 
4. Dehydrate in 95 per cent. and absolute alcohol (1 minute 
in each). 
5. Fast Green. Place 5 drops of a solution of fast green (fast 
green, 1 Gm., absolute alcohol 25 cc., clove oil, 75 cc.) in a watch 
glass of absolute alcohol. Allow the sections to remain in this 
solution until the green color stains the softer tissues properly. 
6. Clear sections in xylol. 
7. Mount in Canada balsam dissolved in xylol. 
8. Label slide, indicating subject and stain combination. 
This method has given excellent results in staining sections of 
a wide variety of fresh and macerated roots, stems and leaves 
which have been cut free hand or on the sliding microtome. 
No. 2. SAFRANIN AND LicHuT GREEN 
1. Stain object with either alcoholic safranin or anilin safranin 
(1 per cent. solution in 50 per cent. alcohol) for 1 to 24 hours or by 
gently warming object in the safranin solution until vapors arise. 
2. Wash out excess of stain with 50 per cent. alcohol. 
3. Dehydrate with 95 per cent. alcohol for two minutes. 
4. Further dehydrate by placing material in absolute alcohol 
for 20 to 30 seconds. ‘ : : 
5. Stain with a saturated solution of light green in clove oil for 
from 3 to 30 minutes. 
6. Clear in xylol for 1 minute. Blot up excess from around 
edge of section. 
