APPENDIX 



I. STAINS AND REAGENTS 



A. Stains. 



1. Iodine solution for staining flagella of euglenae and spermatozoa. 



Make a very strong solution of iodine crystals in 50% alcohol 

 and test for each case, varying strength as necessary. 



2. Methyl green 



Distilled water 100 c.c. 



Methyl green 1 gm. or less 



Glacial acetic acid, a few drops. 



3. Methyl violet 



Distilled water 100.00 c.c. 



Methyl violet 0.05 gm. 



Glacial acetic acid 0.20 c.c. 



This dilute stain is better than a stronger one since it does 

 not readily overstain. The acetic acid is essential, and if 

 an old solution fails to stain the nuclei, the addition of a 

 few drops of the acid will commonly restore its staining 

 power. 



To get the best results in staining blood, substitute for dis- 

 tilled water 0.7% sodium chloride solution for frog blood, 

 and 0.9% for human blood. This prevents swelling and 

 loss of hemoglobin from the red cells, which happens if 

 more dilute solutions are used. 



B. Reagents. 



4. Bela Haller's fluid 



Distilled water 50 c.c. 



Glacial acetic acid 25 c.c. 



Glycerin 25 c.c. 



5. Benedict's solution 



Solution A 



Sodium citrate (NasCeHsOy) 173 gm. 



Sodium carbonate (NaoCOs) 100 gm. 



Dissolve with heat in 600 c.c. distilled water, filter, and make 



up to 850 c.c. with distilled water. 



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