732 THE SPIROCILETE OF SYPHILIS 



1. and 2. Cut the tissues into pieces about 1-2 mm. square, then fix and 

 harden as in the first method. 



3. Wash in distilled water until the pieces fall to the bottom of the vessel. 



4. Leave the pieces of tissue in a ground-glass stoppered bottle for 2-3 

 hours at the temperature of the laboratory then for 3-5 hours at 50 C. 

 in 50 c.c. of the following solution : 



1 per cent, aqueous solution of silver nitrate, - 90 e.c. 



Pyridine, 10 



5. Wash in distilled water. 



6. Immerse for 3 or 4 hours at the temperature of the laboratory in the 

 following solution which must be freshly prepared. 



4 per cent, aqueous solution of pyrogallol, - 90 c.c. 



Pure acetone, - - 10 



Pyridine, - - 17 



7. On taking out of the reducing bath, dehydrate in absolute alcohol, 

 clear in xylol and embed in paraffin. 



8. Cut, stain the sections on the slide with a 2 per cent, aqueous solution 

 of toluidine blue (Buschke and Fischer) and differentiate in absolute alcohol 

 or Unna's ether-glycerin solution. (This step is not essential but by adopting 

 it the groundwork is stained blue and the relationship of the parasites to the 

 cells can be made out.) 



9. Dehydrate in absolute alcohol, oil of bergamot and xylol. Mount in 

 balsam. The treponemata are stained black. 



SECTION III. THE DETECTION AND IDENTIFICATION OF THE 

 TREPONEMA PALLIDUM IN THE TISSUES. 



The distribution of the Treponema pallidum in syphilitic tissues is very 

 irregular. In similar lesions, the parasites are in some cases numerous, in 

 others so few in number that several preparations have to be examined before 

 one or two are seen. Further, a delicate technique has to be adopted, so 

 that several examinations are in most cases essential. 



In the lesions of congenital syphilis the treponemata are far more numerous 

 than in the lesions of acquired syphilis. In the latter case the parasites can 

 most easily be found in the primary lesions and in the secondary papules. 

 Treponemata are rarely seen either in the blood or in tertiary lesions. 



Herscheimer and Opificus advise the routine use of preparations made 

 during the night as they believe that the micro-organism is then present in 

 larger numbers. 



1. Collection of material. 



Chancre. Cleanse the surface of the chancre carefully to expose the dermis. 

 Scrape it gently and repeatedly with a vaccinating lancet in order to produce 

 a slight exudation of serum, a sort of serous dew, and prepare films with the 

 exudate (Thibierge, Ravaut and Le Sourd). 



Alternatively, the chancre may be partially or totally excised and so pro- 

 vide material for cutting sections. This, however, may not always be 

 practicable. 



Rose spots and papules. The simplest method is to scarify the part and 

 collect the exudate. Better, after lightly scarifying, place a small Bier's 

 cupping glass (Zabolotny) or small blister over the lesion and use the serous 

 fluid which collects. 



Gummata. The fluid in softened gummata contains no treponemata. 

 Material from the walls of gummata must be used for the purpose of demon- 

 strating the organism. 



