BILE COMPONENTS 



41 



BISMUTH PIGMENTATION 



chloride for 6 hours ; fix 18 hours in 10% 

 formalin; dehydrate rapidly in alcohol, 

 clear in benzol and embed in paraffin. 

 The bile components, precipitated by 

 barium chloride, can be stained with 

 acid dyes especially the acid fuchsin in 

 Mallory's connective tissue stain (Fors- 

 gren, E., J. Morph., 1929, 47, 519-529). 



Bile Pigments. Histochemical reaction. 

 Fix in 10% formalin or in alcohol. Pro- 

 longed fixation is contraindicated. Fix 

 paraffin sections to slides with egg 

 albumen. Deparaffinize and immerse 

 in 2 or 3 parts Lugol's solution and 1 

 part tincture of iodine, 6-12 hrs. Wash 

 in aq. dest. and cover with sodium hypo- 

 sulphite (5% aq.) 15-30 sec. until de- 

 colorized. Wash in aq. dest. and stain 

 with alum carmine 1-3 hrs. Wash in 

 aq. dest., dehydrate in acetone, clear 

 in xylol and mount in balsam. Bile 

 pigment granules emerald green (Stein, 

 J., C. R. Soc. de Biol., 1935, 120, 1136- 

 1138). See Gmelin's Test. 



Bilharzial Cercariae. For intra - vitam 

 staining examine in serum plus a little 

 neutral red. For permanent prepara- 

 tions fix in hot lactophenol (equal parts 

 lactic acid, carbolic acid, glycerin and 

 aq. dest.). Stain with alcoholic borax- 

 carmine. Mount in following: dissolve 

 by boiling gum tragacanth 3 parts and 

 gum acacia 1 part in aq. dest. 100 parts. 

 Add equal parts lactophenol and use 

 filtrate. (Marshall, A., Lab. J., 1937, 

 7, 565-569). 



Bilirubin, a reddish bile pigment which is 

 isomeric or identical with Hematoidin 

 and which by oxidation can be converted 

 into the green BiliTerdin, see Bile 

 Pigments, tFrobilin and Van den Bergh 

 Test. 



Biliverdin, a green bile pigment produced 

 by oxidation of Bilirubin. See Bile 

 Pigments. 



Bindschedler's Green (CI, 819). A basic 

 indamin dye easily reduced to a sub- 

 stituted diphenylamine. See use as a 

 Redox dye in study of metabolism of 

 tumor tissue (Elliott, K. A. C. and 

 Baker, Z., Biochem. J., 1935, 29 (2), 

 2396-2404). 



Biotin, see Vitamins. 



Bird's Eye Inclusions. Some of these 

 bodies, and the so-called Plimmer's 

 Bodies, seen in cancer cells are ap- 

 parently greatly enlarged Centrosomes. 

 Methods and results are given by Le- 

 Count, E. R., J. Med. Res., 1902, 7 

 (N.S. 2), 383-393. 



Bismark Brown Y (CI, 331) — basic brown, 

 G, GX, or GXP, Excelsior brown, 

 leather brown, Manchester brown, 

 phenylene brown, Vesuvin — A mixture 

 of basic dis-azo dyes of different shades. 

 Quite widely employed, see Blaydes, 



G. W., Stain Techn., 1939, 14, 105-110 



for use with plant tissue. 

 Bismiocymol (see Pappenheimer, A. M. 



and Maechling, E. H., Am. J. Path., 



1934, 10, 577-588. 

 Bismuth. Microchemical detection of: 



1. Method of Christeller-Komaya. 

 Make frozen sections of formalin fixed 

 tissues. A = quinine sulphate, 1 gm.; 

 aq. dest., 50 cc; nitric acid, 10 drops. 

 B = potassium iodide, 2 gm., aq. dest., 

 50 cc. Immediately before use mix 

 equal parts A and B and add 2 drops 

 nitric acid, C.P. After treating sec- 

 tions with this for 1 min. wash very 

 quickly in 10 cc. aq. dest. + 2 drops 

 nitric acid. Mount section on slide. 

 Dry, counterstain with gentian violet. 

 Bismuth appears as dark brown grains 

 (Lison, p. 98). See Komaya, G., Arch, 

 f. Dermat. u. Syph., 1925, 149, 277-291 

 (good colored figures) and Califano, L., 

 Zeit. f. Krebsf., 1927-28, 26, 183-190. 



2. Another modification of the 

 Komaya method is given by Castel, P., 

 Arch. Soc. d. Sci. Med. et. biol. de 

 Montpellier, 1934-35, 16, 453-456 as 

 follows : Dissolve 1 gm. quinine sulphate 

 in 50 cc. aq. dest. with aid of a few drops 

 of sulphuric acid. Dissolve 2 gm. 

 potassium iodide in 50 cc. aq. dest. 

 Mix, apply to section, gives red ppt. 

 of salts of bismuth in form of iodo- 

 bismuthate of quinine or double iodide 

 of bismuth and quinine. See Pappen- 

 heimer and Maechling's (Am. J. Path., 

 1934, 10, 577-588) study of nuclear 

 inclusions in the kidney. 



Bismuth Pigmentation. Histochemical 

 identification as advised by Wachstein, 

 M. and Zak, F. G., Am. J. Path., 1946, 

 22, 603-611 depends on ability of hydro- 

 gen peroxide to decolorize bismuth 

 sulfide instantaneously and of a slightly 

 modified Castel reagent to change bis- 

 muth sulfate into an orange red deposit. 

 Treat deparaffinized, or frozen, sec- 

 tions with few drops superoxol (30% 

 hydrogen peroxide, Merck) from dark 

 bottle kept in refrigerator. In a few 

 seconds black of bismuth sulfide dis- 

 appears. Wash thoroughly in tap wa- 

 ter and place in Coplin jar containing 

 modified Castel reagent made as fol- 

 lows: Dissolve 0.25 gm. brucine sulfate 

 (Merck or Eastman Kodak) in 100 cc. 

 aq. dest. plus 2 or 3 drops concentrated 

 sulfuric acid. Then add 2 gm. potas- 

 sium iodide, keep in a brown bottle and 

 filrer before use. After 1 hr. transfer 

 sections to another jar containing some 

 of reagent diluted with 3 parts aq. dest., 

 and shake gently to remove precipi- 

 tates. Remove most of fluid from sec- 

 tions by blotting and cover with levu- 

 lose solution made by dissolving 30 gm. 

 levulose in 20 cc. aq. dest. at 37 °C. for 



