1V6 



Journal of Applied Microscopy. 



but few of the gonococci, and these may 

 be associated with other germs. 



Gonococcus. 



By means of a looped platinum wire, a 

 portion of the mixture is transferred 

 to a clean cover-glass and spread over as 

 much of the surface as possible. When 

 the specimen has dried in the air it is 

 taken up with a pair of forceps and fixed 

 in the flame. Overheating must be 

 avoided, since it interferes with the 

 proper staining of the germs. Usually, 

 a single passage through the flame will 

 be sufficient to fix the specimen to the 

 cover glass, so that it will not wash off 

 during the subsequent manipulation. 



The fixed specimen, held in the forceps, 

 is then covered with a dilute solution of 

 niethylen blue. In about one-third to 

 one-half minute the dye is washed off, 

 the under side of the cover glass is wiped 

 clean and dry, and finally the cover glass 

 is inverted onto a clean glass slide. A 

 little water should be present between 

 the specimen side of the cover-glass and 

 the glass slide. 



The preparation is now ready to be 

 examined under the microscope. The 

 gonococcus can be seen with the one- 

 sixth or one-eighth-inch objective, but it 

 is advisable for the beginner to employ, 

 if possible, the one-twelfth oil-immersion 

 objective. Even with this lens the 

 organism will appear as very minute 

 points or dots, which may easily be over- 

 looked. The observer will notice, first of 

 all, the large round cells. Within each 

 of these cells there will be usually seen 

 one or two large bodies which are stained 

 a deep blue. These are the nuclei, and 

 the cells themselves are the pus cells. 

 The gonococci are vastly smaller than 

 these pus corpuscles. On careful exami- 

 nation groups of the dot-like gonococcus 

 will be found either between the cells, or 

 in some cases within the pus cells. 



The methylen blue is prepared as fol- 

 lows: A saturated alcoholic solution of 

 the dye is first prepared. This will keep 

 indefinitely and can be kept therefore in 

 stock. One part of this strong solution 

 should be diluted with three to four 

 parts of distilled water. This dilute dye 

 will keep for some time, and it is that 

 which is used for staining purposes. 



Loffler's methylen blue solution is 

 excellent for staining gonococci as well 

 as the diphtheria bacilli. It does not 



deteriorate on keeping. It is prepared by 

 adding 30 cc, of the saturated solution 

 of methylen blue to 100 cc. of a 0.01 per 

 cent, solution of potassium hydrate. 



DIPHTHERIA. 



True diphtheria is due to the Loffler 

 bacillus, whereas certain diphtheria-like 

 conditions of the throat are due to other 

 organisms. Inasmuch as the dangerous 

 form of clinical diphtheria is that due to 

 the Loffler bacillus, and further, since 

 antitoxin is of value only in true diph. 

 theria, it is evident that the recognition 

 of the cause of this disease is essential 

 to its correct diagnosis and treatment. 



The Loffler bacillus is especially present 

 in the false membranes which form on 

 the mucous surfaces of the throat. A 

 portion of the membrane can be removed 

 with a blunt instrument such as a 

 spatula, or better by means of a cotton 

 swab. This is prepared by rolling a tuft 

 of cotton over the end of a stout wire or 

 stick of wood. If the material removed 

 is to be used for growing the bacteria 

 present, it must of course be sterilized 

 before hand. For this purpose the swab 

 is placed in a test-tube, the mouth of 

 which is then plugged with cotton. The 

 tube is then heated in a dry-heat oven 

 at 150 degrees C. for at least one-half 

 hour. 



mv , ««• 



•»il» 





The Loffler Bacillus. 



The diagnosis of diphtheria can be 

 made by the direct examination of the 

 false membrane. It is advisable, how- 

 ever, to supplement the preliminary 

 examination by transplanting some of 

 the material to a tube of sterile blood- 

 serum. The sterile swab should be 

 rubbed firmly over the affected surface 

 and then rubbed over the surface of the 

 serum. The culture tube shoul then be 

 .set aside in a warm place, preferably in 

 an incubating oven which is kept at the 

 temperature of the body. In from 

 eighteen to twenty-four hours numerous 

 colonies develop on the blood serum. 

 Sterile swabs and blood-serum tubes can 

 be purchased on the market and several 

 of the sets should always be kept on 

 hand. 



A portion of the membrane removed 

 from the throat can be rubbed thoroughl> 

 over the surface of a cover-glass. The 

 latter should then be fixed by passing 

 through the fiame, and finally stained 

 with either of the methylen blue 

 solutions mentioned above under 

 " Gonorrhoea." 



