MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



4. Mix equal volumes of solutions A and B and filter at once 

 onto the slides . 



5. Heat the slides over the bunsen flame or on a hot plate until 

 steam rises, but do not allow the stain to boil. 



6. Allow the hot stain to act for three to five minutes ; then pour 

 off excess and allow the slide to cool somewhat before rinsing 

 with distilled water. 



7. Dry thoroughly; then mount in cedarwood immersion oil 

 or in liquid paraffin. 



Results: 



Elementary bodies are stained sharply a light violet colour. 



Note: Instead of methyl violet the following solution may be 

 used, in which case the elementary bodies will stain a pinkish 

 colour: Basic fuchsin 10% in absolute alcohol i volume; distilled 

 water 9 volumes. 



Reference: Gutstein, M. (1937); J', of Path, and Bad., 45, no. i, 313-4. 



METHYLENE BLUE - CARBOL FUCHSESf 

 For flagella and capsules in bacterial films 



Solutions required: 



A. Potash alum, saturated aqueous 40 ml. 

 Tannic acid aqueous 10% . . 40 ml. 

 Methylene blue, saturated in abso- 

 lute alcohol . . . . . . 10 ml. 



Note: This solution deteriorates after a week or 

 ten days. 



B. Carbol fuchsin (Ziehl Neelsen) . . 5 ml. 

 Distilled water . . . . • • 45 n^l- 



Technique: 



1. Smears are made on scrupulously clean slides by spreading a 

 minute speck of the culture with distilled water. 



2. Allow to dry and fix by flaming in the usual manner. 



384 



