CAPSULE STAIN 



57 



CARBOL-THIONIN 



fastness 3. 0.1 gm. in 95 cc. aq. dost. 

 + 5 cc. 5% aq. ammonium alum + 0.5 

 cc. acetic acid stains plant tissues blue 

 or black. Can be employed in prefer- 

 ence to Cyanine. Should stain well 

 after Erythrosin (Eniig, p. 58). 

 Capsule stain. 1. Hiss' method for smears 

 (McClung, p. 145). Dry organisms in 

 ascitic or serum medium on slides. 

 Stain, slightly heated in 5-10 cc. satu- 

 rated ale. gentian violet or basic fuchsin 

 made up to 100 cc. aq. dest., few sec. 

 Wash off dye with 20% aq. copper sul- 

 phate crystals. Dry by blotting. See 

 also: Huntoon, F. M., J. Bact., 1917, 2, 

 241. See Pasteurella. 



2. W. H. Smith's method for sections 

 (Mallory, p. 275). Cover deparaffin- 

 ized sections of Zenker fixed material 

 with Anilin Crystal Violet (either 

 Ehrlich's or Stirling's). During few 

 seconds warm by passing slide through 

 flame 2 or 3 times. Wash in Gram's 

 Iodine solution followed by formalin 

 (commercial). Decolorize in 95% ale. 

 Quickly wash again in Gram's iodine. 

 Cover with aniline green eosin and heat 

 as before. To make this shake 1 part 

 aniline green with 200 parts 3-6% aq. 

 eosin yellowish W.S. and after 1-2 hrs. 

 remove ppt. bj' filtering. Wash in aq. 

 dest. Dehydrate in 95% and abs. ale, 

 clear in xylol and mount in balsam. 

 Bacterial capsules, red; Gram positive 

 bacteria, blue. Mallory says that a 

 stronger iodine may be desirable (iodine, 

 1 gm., potassium iodide, 2gm. ;aq. dest., 

 100 cc.) and that the times must be 

 suited to each preparation. 



3. Churchman's (S. Bayne-Jones in 

 Simmons and Gentzkow, p. 385). 

 Flood air-dried films with Wright's 

 stain and leave until almost evaporated 

 to dryness. Original blue of stain is 

 replaced by pinkish color. Wash 

 quickly in water, or in Clark and Lubs 

 buffer pH 6.4-6.5. Do not blot but dry 

 with fan. Body of organisms, blue; 

 capsular material, purplish-pink; often 

 surrounded by capsular membrane or 

 peripheral zone, deep purplish-pink. 



Capsule Substance. This obviously is un- 

 der investigation in many sorts of cells 

 and the methods introduced for one 

 kind may well be of service for others. 

 See Cell Membrane for physical proper- 

 ties, thickness, etc. See Adhesiveness 

 and Acid Fast Bacilli. Under Gram 

 Stains is a description of the mechanism 

 of their action which includes data ob- 

 tained by use of the enzyme, ribonu- 

 clease, on the nature of walls of Gram 

 positive bacteria. Under Enzymes, see 

 enzymatic destruction of capsules of 

 pneumococci. 



Carbanthrene Blue GO (CI, 1113), Carban- 



threne Brilliant Orange RK, Carban- 

 threne Jade Green (CI, 1101), Carban- 

 threne Red BN (CI, 1162) Carbanthrene 

 Red BN (CI, 1162) and Carbanthrene 

 Violet 2R (CI, 1104) all of NAC are 

 referred to by Emig, p. 64. 



Carbohydrates, see Starch. 



Carbol-Anilin Fuchsin methylene blue 

 method for Negri bodies (Goodpasture, 

 E. W., Am. J. Path., 1925, 1, 547-582). 

 Fix in Zenker's fluid 24 hrs. (not Helly's 

 fluid). Color for 10-30 min. in mixture 

 made by adding 1 cc. of pure phenol and 

 1 cc. of anilin oil to 100 cc. of stock 0.5% 

 basic fuchsin in 20% alcohol. Wash in 

 running water, blot with filter paper and 

 decolorize with 95% alcohol until sec- 

 tions become pink. Then wash in water 

 and stain with Loeffler's methylene 

 blue, 15-60 sec. Wash again in water. 

 Dehydrate and destain for few sec. in 

 absolute alcohol, clear in xylol and 

 mount in balsam. Negri bodies, crim- 

 son; background, blue. Also excellent 

 for Borrel Bodies. 



Carbol-Crystal Violet. Because the solu- 

 tions as prescribed in Nicolle's original 

 formula for carbol gentian violet tend 

 to gelatinize, the following formula is 

 recommended by Conn, H. J., Stain 

 Techn., 1946, 21, 31-32: Mix solution of 

 0.4 gm. crystal violet C. C. in 10 cc. 

 95% ethyl alcohol with solution of 1 gm. 

 phenol in 100 cc. aq. dest. 



Carbol-Fuchsin, The original formula of 

 Ziehl has been much modified. Ziehl- 

 Neelsen is sat. ale. basic fuchsin, 10 cc. ; 

 5% aq. carbolic acid, 90 cc. Verhoeff 

 (F. H., J.A.M.A., 1912, 58, 1355) advises 

 basic fuchsin, 2 gm.; abs. ale, 50 cc; 

 melted carbolic acid crystals, 25 cc. 

 McClung (p. 136) suggests mixing 10 cc. 

 3% basic fuchsin (90% dye content) 

 with 95 cc. 5% aq. phenol. The im- 

 portant thing is the character of the 

 fuchsin not its concentration relative to 

 carbolic acid. Carbol-fuchsin is em- 

 ployed in stains for Acid Fast Bacilli. 

 DeipoUi, G. and Pomerri, G., Mon. 

 Zool. Ital., 1938, 49, 123-124 have ad- 

 vised its use as follows for Nissl Bodies. 

 Fix small pieces in 95-98% alcohol or in 

 10% formalin water or in physiological 

 saline 24 hrs. or longer. Stain deparaf- 

 finized sections 3-4 min. in carbol- 

 fuchsin (basic fuchsin, 0.2 gm.; cone, 

 phenol, 1 cc; 95% ale, 2 cc. ; aq. dest. 

 20 cc.) 2.5 cc. ; aq. dest., 100 cc. ; glacial 

 acetic acid, 0.5 cc. Wash rapidly in aq. 

 dest. and destain in: aq. dest., 100 cc; 

 formalin, 1 cc. ; glacial acetic acid, 1 cc. 

 Wash in aq. dest., dehydrate in alcohols, 

 clear in xylol and mount in neutral 

 balsam. Nissl bodies and nucleoli dark 

 red, rest unstained. 



Carbol-Thionin, see King's. 



