FLAGELLA STATXS. 



191 



Fischer's Flai;clla Slain. 



This is a slight modification of Dr. Corner's, 

 wliicli is itself a modification of T^oefflers. 

 ( I ) INfordant ; 



I )ry tannin 2 



A\'ater 20 



Solntion ferrons snlphatc { r : 2) . . . . 4. 

 Cnncentrated alcoholic solntion hasic 



fnchsin i 



Filter This mordant will keep for several 

 weeks. Drop it on the co\-er and heat over a 

 gentle flame nntil steam rises ; then continue one- 

 half minute longer without hoiling. Wash. 

 Treatment with alcohol is unnecessary. 



(_>) Stain: Place on the cover a few drrjps of 

 concentrated watery solution of hasic fuchsin. 

 II eat slowly, so that steam rises after aliout I 

 minute. Then expose for one-half minute 

 longer, so that the stain hods up once or twice. 

 Wash, dry, and mount. Fischer sa} s tannin ah- 

 sorhs moisture readily, and advises keeping it 

 in a desiccator. 



Biiiigc's Flagclla Staiii. 

 ( I ) Mordant ; 



Tannin 20, water 100 30 



Liquor ferri sescpiichlorati i 



Water 20 



Saturated watery solution hasic 



fuchsin 5 



This must ripen some weeks hy exposure to 

 the ;iir in a tfask loose]}- plugged with cotton. 

 (2) Stain: Carliol-fuchsin. 



Expose to the filtered mordant 3 minutes, 

 using heat if necessary. Wash and stain. 



I'an Ei-ii!c}igciit's F'lagclla Shiiii. 

 This is made as follows ■ 

 ( [ ) Mordant : 

 ( )snuc acid (3 per cent water solu- 

 tion") SO 



Tannin ( 10 to 25 per cent in water ) . . 100 

 Four drops of glacial acetic acid may be added 

 to this. 



(2) Silver hath: 0,25 to 0.5 jier cent nitrate of 

 silver dissolved m distilled w.ater in a very clean 

 hottle. 



(3) Reducing and strengthening hath: 



Gallic acid 5 



Tannin 3 



Fused sodium acetate ( some hooks 



say fused potassumi acetate) lo 



Distilled water 350 



The flamed cover-glass (it may he unflamed) 

 is lirst covered with the mordant for one-half 



hour, or if in a thermostat at 50° C. for 5 or 10 

 minutes. The mordant is then carefully re- 

 moved hy thorough washing in water, alcohol 

 (some say ahsolute alcohol), and water. The 

 cover (film side up) is now put into the silver 

 bath (a few cuhic centimeters in a small, per- 

 fectly clean heaker or watch-glass) for a few 

 seconds, during which it is gently agitated. 

 Without rinsing, it is put next into a few culiic 

 centimeters of the reducing solution and gently 

 a.gitated until the fluid begins to blacken. It is 

 then Haslie<l in water and examined. If not 

 stained deeply enough the cover is returned to 

 the silver hath, then once more passed through 

 the reducing" hath. It is finally dried and 

 mounted in h.als.ani All the dishes must be 

 scrupulousl}' clean. The fluids must not be con- 

 taminated by the fin.gers nor hy dipping iron or 

 steel instruments into them. 



Kuntze has suggested some improvements. 

 (See Centralh f. Bakt., I Abt., Bd. XXXII, 

 1902, pp. 555-560.) 



FitncliVs Flagclla Stain. 



(i) Mordant: 



Tannic acid, 10 per cent acj. sol 10 



Corrosive sublimate, sat. aq. sol 5 



Alum, sat. a([. sol 5 



Carbol-fuchsin 5 



Fleat on co\er until steam rises; keep at this 

 temperature i minute; then wash, dry, and 

 stain. 



(2) Stain; 



.Alum, sat. ( cold ) aq. sol 10 



(.jenti:m violet, sat. ale. sol 2 



Kendall, in Journ. Applied Micr., Vol. V, 

 1902, p 1X3(1, says this has proved a very satis- 

 factory stain to himself and his associates. 



Another formula is given as follows: 

 (I) Saturated (cold) aqueous solution c)f 



alum 10 



Saturateil alcoholic solution of gentian 



violet I 



( 2 ) Tannic acid ( tannin ) i 



Distilled water 10 



A mixture of these two fluids is put on the 

 flamed cover, which is held over the flame and 

 gently heated until nearly ready to boil. The 

 cover is then put aside for i minute, after which 

 it is washed in water, dried, and mounted in bal- 

 sam. If the mixed mordant-stain is filtered be- 

 fore using, it is best to stain a second time for a 

 moment in anilin-water gentian violet. 



