TO STAIN PROTOZOA IN SECTIONS 125 



Pyrogallic acid 4 grammes 



Acetone 10 c.c. 



Pyridin puriss 15 grammes 



Distilled water 100 c.c. 



8. Wash well in water. 



Take through alcohols of increasing strength up to 

 absolute, keeping in each strength for twenty-four 

 hours. 



9. Clear, embed, cut very thin sections, mount, 

 remove paraffin, again clear and mount in xylol balsam. 



The spirochaetes if present are black and show up 

 against the pale yellow color of the background. 



Weak carbol fuchsin, neutral red or toluidin blue 

 can also be used to stain the background if desired, 

 after the removal of the paraffin in step 9. 



To Demonstrate Protozoa in Sections (Leishman) . 



Reagents required : 



Leishman' s Polychrome stain. 

 Acetic acid i in 1500 aqueous solution. 

 Caustic soda i in 7000 aqueous solution. 

 Distilled water. 



1. Mount section, remove paraffin and take into 

 distilled water as usual (vide page 121). 



2. Drain off the excess of water. 



3. Cover the section with diluted Leishman (i part 

 stain, 2 parts distilled water) and allow to act for five 

 to ten minutes (until tissue appears a deep blue) . 



4. Decolourise with acetic acid solution until only 

 the nuclei appear blue (examine the section wet, with 

 low power objective) . 



5. If the eosin colour is too well marked treat with 

 the caustic soda solution until the desired tint is ob- 

 tained (as seen with the J-inch objective). 



6. Wash with distilled water. 



7. Rapidly dehydrate with alcohol. 



8. Clear with xylol. 



9. Mount in xylol balsam. 



