174 ATLAS OF BACTERIOLOGY. 



Bunge lias recently employed a somewhat different 

 method which also gave us good results, but, like 

 Loffler's method, occasionally left us capriciously in 

 the lurch. 



(1) Preparation of the specimen, according to 

 Loffler. 



(2) Heating with Bunge' s mordant for one minute 

 until steam is produced. 



(3) Careful cleaning with water and drying. 



(4) Warming slightly with carbolized gentian violet 

 or carbolized fuchsin. 



(5) Washing in water, drying, and mounting in 

 Canada balsam. 



Most of our specimens are prepared with Bunge's 

 mordant which is several months old. 

 5. Staining of Endospores.* 

 According to Hauser : 



(1) Preparation of the specimen. (It should be 

 drawn ten times, instead of three times, rapidly 

 through the flame.) 



(2) Staining with watery fuchsin or carbolized 

 fuchsin (Ziehl's solution) ; the preparation, over the 

 flame, is covered freely with the staining fluid, and 

 heated (not boiled) one to two minutes until there is 

 an indication of simmering. The evaporating stain- 

 ing fluid is replaced constantly by fresh fluid. 



(3) Washing with acid alcohol, \ until the red color 

 of the preparation is almost gone. 



* Arthrospores possess no undisputed color reactions. For 

 metachromatic corpuscles, Ernst's and Bunge's granules, pre- 

 liminary stages of spores, vide page 71. 



f Instead of acid alcohol we may also use thirty per cent 

 nitric acid, rive or twenty five per cent sulphuric acid, but 

 these must be allowed to act for a shorter period. 



