BACTERIOLOGICAL TECHNIC IOI 



Bacteria not Stained by the Gram Method 



Diplococcus of meningitis (intracellular) . 



Diplococcus of gonorrhea. 



Micrococcus melitensis. 



Bacillus of chancroids (Ducrey's). 



Bacillus of dysentery (Shiga's). 



Bacillus of typhoid fever. 



Bacillus of bubonic plague. 



Bacillus of influenza. 



Bacillus coli communis. 



Bacillus pyocyaneus. 



Bacillus of Friedlander. 



Bacillus proteus. 



Bacillus mallei (glanders). 



Spirillum of Asiatic cholera. 



Spirillum of relapsing fever. 



5. Pappenheim's Stain 



Sat. Aqueous Sol. Methyl Green, 50 cc. 



Sat. Aqueous Sol. Pyronine, 15 cc. 



Mix and filter. This is much used for staining bacteria in pus and 

 other pathological secretions. The bacteria are stained a bright red, while 

 the cell nuclei are blue to purple. 



6. Smith's Stain 



Stock Sol. Basic Fuchsin, 10 cc. 

 Methyl Alcohol, 



Formaldehyde, each 10 cc. 



Distilled Water, to make 100 cc. 



Mix and filter. Let stand for twenty-four hours before using. Renew 

 in three weeks. The stain is much used to distinguish between bacteria 

 and nuclear substances. Allow the stain to act for from two to ten 

 minutes. 



7. Flagella Staining 



Care is necessary in staining flagellse. Numerous methods have been 

 recommended, but Pitfield's method, -as modified by Muir, is perhaps the 

 best and at the same time comparatively simple. The following solutions 

 are required: 



a. Mordant 



Tannic Acid (10 per cent. Aq. Sol.), 10 cc. 



Sat. Aq. Sol. Mercuric Chlor., 5 cc. 



Sat. Aq. Sol. Alum, 5 cc. 



Carbol-Fuchsin, 5 cc. 



Mix, shake, filter or centrifuge. This soluton does not keep longer 

 than one week. 



