384 Special Procedures II (chap. 23) 



W^orking solution: 



carmalum stock 5.0 ml. 



glacial acetic acid 0.4 ml. 



distilled water 100.0 ml. 



procedure: stain for 48 hours, no destaining. Dehydrate as above. 

 Carmalum is good and requires no destaining. 



Hematein (kornhauser, 1930) 



solutions: 



Stock solution: 



Grind 0.5 gm. hematein with 10 ml. 95% alcohol and add to 5% 

 aqueous aluminum sulfate. 



Working solution: 



Dilute above 1: 10 with distilled water. 



procedure: 



1. Stain overnight. 



2. Transfer to 70% alcohol. 



3. Destain in acid alcohol, 5% HCl in 70%, alcohol. 



4. Blue in alkaline alcohol, ammonia or sodium bicarbonate in 70% 

 alcohol. 



5. Dehydrate and mount as above. 



Hematein is good for flatworms. 



Alum cochineal used to be popular but has become largely replaced by 



carmine stains. 



Alum hematoxylin may be used for small organisms if not too dense. 



Celestin blue method, Demke (1952) may be used. 



Protozoa 



fixation: hot Schaudinn's, 50°C: 5-15 minutes. Other fixatives are 

 formol Bouin's, acetic, or mercuric chloride acetic. 



Organisms such as Paramecium should be concentrated by centri- 

 fuging. Quickly pour off some of the solution and add fixative. 

 Amoeba will settle on the bottom of a clean culture dish. Decant 

 off most of the culture media, leaving the bottom barely covered. 

 Quickly pour hot Schaudinn's or Bouin's (50-60°C) over the organ- 

 isms. After a few minutes add an equal amount of 85% alcohol. Care- 

 fully loosen any amoebae clinging to the bottom and collect the solu- 

 tion in a centrifuge tube. For both Paramecium and Amoeba, allow 



