BILE CAPILLARIES 



36 



BIOLOGICAL STANDARDS 



Pus cells (neutrophiles), scattered 

 through the specimen and bile stained, 

 which, when occurring in fair numbers, 

 indicate cholecystitis. Unstained pus 

 cells associated with mucus are generally 

 from the mouth. (2) Bile colored epi- 

 thelial cells and cellular debris suggest 

 chronic cholecystitis. (3) Cholesterin 

 crystals are identifiable as opaque or 

 translucent, flat, rhombic plates or 

 irregular masses. (4) Large amounts of 

 light brown granules or dark black- 

 brown ppt. of calcium bilirubinate are 

 suggestive of gall stones. (5) Tiny gall 

 stones (bile sand) are identifiable by 

 their concentric lamination. Negative 

 findings are not, he is careful to point 

 out, conclusive of absence of lesions. 

 Bile Capillaries. 1. Hematoxylin staining. 

 Clara, M., Zeit. f. mikr. Anat. Forsch., 

 1934, 35, 1-56 advises treatment of cel- 

 loidin sections of pieces of liver fixed in 

 Alcohol Formalin, formalin — absolute 

 alcohol — acetic acid (20:80:1) and other 

 mixtures by the Stolzner Holmer tech- 

 nique and his own method. According 

 to the former, mordant the sections in 

 liquor ferri sesquichlorati (try 10% 

 aq. ferric chloride) 30-45 min. Wash 

 quickly in aq. dest. Stain in ripe 0.5% 

 aq. hematoxylin, 20-30 min. Wash 

 quickly in water. Differentiate in much 

 diluted liquor ferri sesquichlorati. 

 Wash again quickly in water. Blue 

 with dilute aq. lithium carbonate. 

 Wash in spring water (tap water will do ) . 

 Dehydrate, clear and mount. According 

 to Clara, mordant the sections in equal 

 parts A and B at 40-50 °C. for 24 hrs. 

 (A = potassium bichromate, 2.0 gm. ; 

 chrome alum, 1 gm., aq. dest., 30 cc. 

 B = ammonium molybdate, 2.5 gm.; 

 chromic acid, 0.25 gm.;aq. dest., 100 cc.) 

 Wash briefly in aq. dest. Stain in 

 Kultschitzky's Hematoxylin. Wash in 

 spring water. Dehydrate, clear and 

 mount in balsam. See Clara's illustra- 

 tions. 



2. Rio Hortega silver carbonate method 

 adapted by Mclndoe, A. H., Arch. 

 Path., 1928, 6, 598-614. Fix small pieces 

 normal human liver at least 20 days in 

 10% formalin. Heat gently but do not 

 boil and cool several times thin frozen 

 sections for 20 min. in silver bath until 

 they are uniformly of a golden brown 

 color. (To make the bath combine 30 

 cc. 10% aq. silver nitrate and 10 cc. sat. 

 aq. lithium carbonate. Wash ppt. re- 

 peatedly with doubly distilled water, 

 decanting washings. Add 100 cc. doubly 

 distilled water to ppt. Dissolve ^i 

 of it by adding ammonia water drop by 

 drop. Filter supernatant fluid into 

 opaque bottle and store in dark where it 

 can be kept 2-4 weeks. For use take 



5 cc. of this stock solution and add 5 cc. 

 aq. dest. and 2-3 drops pyridine.) 

 Wash quickly in aq. dest. Place in 20% 

 neutral formalin, 1 min. Fix in 2% aq. 

 sodium thiosulphate, ^-1 min. Wash 

 thoroughly in tap water, 2-3 days adding 

 a little neutral formalin. Dehydrate 

 in 95% and abs. ale, clear in carbol- 

 xylol and mount in balsam. Canaliculi, 

 black. 



Bile Components in hepatic cells. Place 

 small pieces of liver in 3% aq. barium 

 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 Biliverdin, see Bile 

 Pigments, Urobilin 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). 



Binnennetz, see Golgi Apparatus. 



Biological Standards. Vitamins, antitoxins, 



