392 



LABORATORY DIAGNOSIS OF PROTOZOAN INFECTIONS 



Allow them to stand for about an hour, 

 and then skim the foam from the top and 

 pour the remaining liquid into a graduated 

 cylinder. Pour in an equal amount of 

 glycerol, and add 1 g of sodium salicylate 

 for each 100 ml of the mixture. Shake 

 thoroughly and filter thru paper into a 

 clean bottle. Filtration will require 1 to 

 several weeks. It may be accelerated 

 somewhat by pouring a rather small amount 

 at a time into the filter and replacing the 

 paper every few days. Keep a small quan- 

 tity in a vial or bottle provided with a glass 

 rod stuck into the cork and projecting into 

 the albumen. A drop can easily be placed 

 on a slide with this rod. j 



Hollande's Fixative 



Picric acid 4 g 



Copper acetate 2. 5 g 



Formalin 10 ml 



Acetic Qcid 1.5 ml 



Distilled water 100 ml 



Schaudinn's Fixative 



Saturated aqueous mercuric chloride . 66 ml 

 95% alcohol 33 ml 



2. Cool to 50° C. 



3. Filter. 



4. Add 20 ml 1 N HCl to the filtrate. 



5. Cool to 25° C. 



6. Add 1 g dried sodium bisulfite 

 (NaHSOs); this liberates suLfurous 

 acid. 



7. Allow to stand at room temperature 

 24 hours until decolorized. 



8. Store in the refrigerator in small 

 glass-stoppered bottles filled to 

 the top to exclude air. The solu- 

 tion will keep several weeks. It 

 should be straw-colored; if it is 

 red, it should be discarded. 



S0rensen's Phosphate Buffers 



To make M/15 Na2HP04 solution, 

 dissolve 9. 5 g anhydrous Na2HP04 or 11.9 

 g Na2HP04 • 2 H2O in 1 liter distilled water. 

 To make M/15 KH2PO4, dissolve 9.08 g 

 KH2PO4 in 1 liter distilled water. Store 

 separately in pyrex, glass-stoppered 

 bottles. 



To prepare buffered water for the 

 Giemsa stain, mix the following amounts 

 of the solutions (in ml): 



Add 5% acetic acid immediately before 

 use. 



Iron Alum Solution 



Ferric ammonium sulfate (violet crystals 



only) 2 g 



Distilled water 100 ml 



Filter immediately before use. 

 Heidenhain's Hematoxylin (Stock Solution ) 



Hematoxylin 10 g 



100% alcohol 100 ml 



Allow to remain 1 month in a loosely stop- 

 pered bottle before use. To make the 

 staining solution, add 0. 5 ml of the stock 

 solution to 9. 5 ml of distilled water. 



pH6.8 pH7.0 pH7.2 



1.0 M K^HPO^ (174. 180 g KjHPO^ 



in 1000 ml distilled water) .,._.. 4.3 parts 

 1.0 M KH2PO4 (136.092 g KH2PO4 



in 1000 ml distilled water) 0. 7 ports 



This is the stock solution. To pre- 

 pare the final solution used in Balamuth's 

 medium, add 14 parts of distilled water 

 to 1 part of the stock solution. 



Ringer's Solution 



Feulgen Stain 



1. Dissolve 1 g basic fuchsin (certified 

 as suitable for the Feulgen stain) 

 in 200 ml boiling distilled water. 



NaCl 

 NaHC03 

 CaCl2 . 2H2O 

 KCl 



NaHjPO^ • 



HjO 



Distilled water 



6.5g 

 0.2g 

 0. 16g 

 0.14g 

 0.011 g 

 1000 ml 



