BEST'S CAHMINE 



40 



BILE COMPONENTS 



1 gm.; potassium chloride, 5 gm.; aq. 

 dest., 60 cc. Boil gently until color 

 darkens, cool and add 20 cc. cone, am- 

 monia. Allow to ripen 24 hrs. This 

 is stock solution. Used to stain Glyco- 

 gen. See Bensley, C. M., Stain Tech., 

 1939, 14, 47-52. 



Beyer Brown, a diazo dye, stains in aq. or 

 alcoholic solution like a good Ehrlich's 

 hematoxylin (H. G. Cannan, J. Roy. 

 Micr. Soc, 1941,61,88-94). 



Bichromate-Chromic-Osmic mixture, see 

 Champy's Fixative. 



Biebrich Scarlet, water soluble (CI, 280) — 

 croceine scarlet, double scarlet BSF, 

 Ponceau B, scarlet B or EC — An acid 

 dis-azo dye much used in histology. 

 See Bowie. 



Biebrich Scarlet and Picro-Anilin Blue, 

 as a differential stain for connective 

 tissue and muscle (Lillie, R. D., Arch. 

 Path., 1940, 29, 705). Deparaffinize 

 sections of material fixed in formalin, 

 Zenker's or Orth's fluid. Stain for 5 

 min. in following: Dissolve 1 gm. 

 hematoxylin in 95% ale. and 4 cc. 29% 

 aq. FeCls in 95 cc. aq. dest. + 1 cc. 

 cone, hydrochloric acid. Mix and use 

 while fairly fresh. Wash in tap water. 

 Stain for 4 min. in 0.2 gm. Biebrich 

 scarlet + 100 cc. 1% aq. acetic acid. 

 Rinse again in aq. dest. Stain for 4 min. 

 in 0.1 gm. anilin blue W.S. (CC.) + 

 100 cc. sat. aq. picric acid. Wash for 

 3 min. in l%aq. acetic acid. Dehydrate 

 in acetone or alcohol. Clear and mount 

 in salicylic acid balsam. Connective 

 tissue, glomerular basement membrane 

 and reticulum, deep blue; muscle and 

 plasma, pink; erythrocytes, scarlet. 

 (Checked by R. D. Lillie, National In- 

 stitute of Health, Bethesda, Md., April 

 22, 1946.) 



Bielchowsky Silver Methods. These are 

 designed for the nervous system and 

 consist essentially of formalin fixation, 

 silver impregnation, washing, treating 

 with ammoniacal silver solution, wash- 

 ing and reduction in formalin. Several 

 useful modifications are detailed by 

 Addison (McClung, pp. 463-466). See 

 Nervous System, Silver Methods. 



Bile. This frequently comes in for micro- 

 scopic examination of centrifuged sedi- 

 ment. Stitt (p. 761) says that one must 

 be on the lookout especially for: (1) 

 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 ^-f 

 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, h-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 



