TRACHEA 



251 



TREPONEMA PALLIDUM 



to demonstrate secretion of Mucus 

 are detailed by Florey, H., Carleton, 

 H. M. and Wells, A. Q., Brit. J. Exper. 

 Path., 1932, 13, 269-284. Techniques 

 for Nerve Endings are given under this 

 heading but it would be helpful to con- 

 sult Larsell, O. and Dow, R. S., Am. J. 

 Anat., 1933, 52, 125-146 who illustrates 

 what one may expect to find. Tech- 

 niques for Cilia require no special 

 adaptation. Celloidin sections are 

 smoother than paraffin ones. 



Tracer Techniques, see Radioactive Isotopes. 



Trachoma Bodies. These are easily colored 

 by Giemsa's stain. For demonstration 

 of glycogen in them and other pertinent 

 data see Thygeson, P., Am. J. Path., 

 1938, 14, 455-462. 



Evolution forms of Rickettsia tra- 

 chomatis. Fix smears in iodine alcohol, 

 4-5 min. Stain in May-Grimwald, 1 

 part; Giemsa, 1 part; neutral aq. dest. 

 10 parts for 1 hr. Differentiate in 95% 

 alcohol (Foley, H. and Parrot, L., Arch. 

 Inst. Pasteur d'Alg^rie, 1938, 16, 283- 

 292) . See colored plates by the authors. 



Transplantation. This technique provides 

 opportunities for important microscopic 

 studies. See Tooth Germs, Anterior 

 Chamber of Eye. 



Trematodes. Make up stain by mixing 

 1 gm. of dried residue on filter paper 

 from Schneider's aceto-carmine with 

 10 gm. ammonia alum in 200 cc. aq. 

 dest. with aid of heat. When dissolved, 

 cool, filter and to filtrate add crystal 

 of thymol. After fixation bring worms 

 to water or to 20% alcohol. Stain 12-36 

 hrs. depending on size. Remove to 

 water 2 changes. Dehydrate through 

 20, 35, and 50 to 70% alcohol. Place 

 few crystals potassium chlorate in small 

 glass covered dish; add few drops cone. 

 HCl. When chlorine is given off fill 

 dish with 70% alcohol. If deeply 

 stained differentiate in this chlorinated 

 alcohol. If not or the specimens are 

 small ones add it to the alcohol covering 

 them and agitate. When sufficiently 

 destained remove to fresh 80% alcohol. 

 Dehydrate in alcohol. Add cedar wood 

 oil to the absolute until mixture is one 

 half oil. Clear in cedar oil and mount 

 in balsam (Gower, W. Carl, Stain 

 Techn., 1939, 14, 31-32). 



Treponema Pallidum. The organisms can 

 best be seen in the primary lesions by 

 Darkfield examination. The same 

 method is useful for skin and lymph 

 nodes in the secondary stage but for 

 the tertiary lesions in deep lying tissues 

 sections are desirable supplemented 

 by smears. A negative finding is com- 

 forting but does not necessarily signifj' 

 absence of parasites unless confirmed 

 serologically. 



1. Low surface tension stain for 

 smears (Haire, R. D., J. Lab. & Clin. 

 Med., 1938, 23, 1215-1216). Mix 1 gm. 

 Gentian violet (or crystal violet) in 

 mortar slowly adding 100 cc. hexylre- 

 sorcinol. Filter and store filtrate in 

 stock bottle. Stain smears 30 min. 

 Wash in water, dry and examine. Stain 

 on slide must not be heated. Trep- 

 onemas, light purple. 



2. Wright's stain for smears (Mallory, 

 p. 289). To make stain add 1 cc. 

 Wright's stain and 1 cc. 1% aq. potas- 

 sium carbonate to 10 cc. aq. dest. in 

 test tube and heat to boiling. Spread 

 material thinly on cover glass (not slide) 

 and hold level with forceps. Cover 

 with hot stain 3-4 min. After fluid has 

 turned violet, and a yellow metallic 

 scum has formed over it, pour off and 

 repeat process twice with hot stain. 

 Wash in water, dry and mount in balsam. 

 Treponemas, intensely violet. 



3. Giemsa's stain for smears (Giemsa, 

 G., Deut. m-ed. Wochn., 1909, 35, 1751- 

 1752) after Mallory (p. 290). Fix 

 smears for 15 min. in absolute alcohol 

 or pass them through flame thrice. 

 Pour on freshly diluted stain (1 cc. aq. 

 dest. + 1 drop stock Giemsa). Steam 

 gently and leave 15 sec. Decant and 

 add immediately fresh diluted stain, 

 warm and let cool 15 sec. Repeat 4 

 times leaving 1 min. last time. Rinse 

 quickly in running water. Blot. 

 Mount in balsam. Treponemas, dark 

 red. 



4. Fontana-Tribondeau silver method 

 for serum (Fontana, A., Dermat. Zeits., 

 1925-26, 46, 291-293) after Mallory 

 (p. 291). To make silver solution add 

 ammonia water (diluted 1:20) drop by 

 drop to 50-100 cc. 1% aq. silver nitrate 

 until a coffee colored clouding takes 

 place. Air dry thin smears of serum. 

 Pour on few drops Ruge's sol. (aq. dest., 

 100 CO.; glacial acetic, 1 cc; formalin, 

 2 cc.) and change several times during 

 1 min. Rinse in running water. Mor- 

 dant witha little aq. dest., 100 cc; tan- 

 nicacid, 5gm.; liquid carbolicacid, 1 cc. 

 for 20 sec. warming to steaming. Rinse 

 in aq. dest. Treat with silver solution 

 30 sec. heating slightly. Wash in tap 

 water. Dry in air. Mount in balsam. 

 Treponemas, brown to deep black. 



5. Burri's India Ink method for lesion 

 fluid (Mallory, p. 291). Make 1:4 

 suspension of India ink in aq. dest. 

 Sterilize in autoclave, 15 min. i\Iix 

 this in equal parts with fluid from lesion 

 on slide with platinum loop. Spread 

 thinly. Dry and examine. Trep- 

 onema (and bacteria if present), white 

 in brown to black background. 



6. Quick method for demonstration 



