STAINS 



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STAINS 



preparations must be kept in a cool place. When the 

 hardening is completed the organs, after washing in 

 water, are preserved in I r c bichromate solution. The 

 method is particularly recommended for the hardening 

 of organs in a state of postmortem softening. Bie- 

 brich Scarlet {Biebricher Scharlach), a plasma stain. 

 Bielschowsky-Plien Method for staining NissI 

 bodies : Harden the tissue in alcohol or formalin, em- 

 bed in paraffin celloidin, and stain for 24 hours in very 

 dilute kresyl-violet R R (IO drops of a saturated 

 aqueous solution to 50 c.c. of water) ; rinse, dehydrate 

 in the series of ascending alcohols, clear in oil of caje- 

 put ; xylol and balsam. Bignami's Method for the 

 study of the malarial parasite in tissue sections: Fix 

 the tissue for several hours in a solution of sublimate I 

 gm., sodium chlorid 0.75 gm. , acetic acid I c.c, and 

 water 100 c.c. Transfer to alcohol containing tincture 

 of iodin and then to absolute alcohol. Stain in ma- 

 genta Griibler) dissolved to saturation in water or in 

 5 % carbolic acid and transfer to absolute alcohol. The 

 sections may be double stained in a mixture of magenta 

 and aurantia in saturated alcoholic solution. Birch- 

 Hirschfeld Method for amyloid tissues : Stain the 

 sections for 5 minutes in 2% solution of bismarck- 

 brown in 40% alcohol; rinse in absolute alcohol; 

 wash for 10 minutes in distilled water; stain for 5 or 

 IO minutes in 2% gentian-violet solution; wash in 

 water acidulated with acetic acid (10 drops to a watch- 

 glassful of water) ; mount in levulose. Bleu Lu- 

 miare, B. de Lyon, B. de Nuit. See Spirit-blue 

 under Pigments (Illus. Diet). Boeck's Method 

 for preparations of epiphytic bacteria: Extract the 

 oil by alcohol and ether; stain for from 30 to 60 

 seconds with Sahli's methylene-blue ; transfer to water 

 containing a fragment of resorcin, and after a minute 

 to alcohol for an hour ; decolor in a weak solution of 

 hydrogen dioxid (if necessary), and dehydrate, clear, 

 and mount in the usual way. Bohm's Method. 

 I. For the demonstration of excretory capillaries. 

 Treat very small cubes of tissue for 3 days in a mix- 

 ture of 4 volumes of 3 ^ potassium bichromate and I 

 volume of I % osmic acid ; then for from 24 to 48 

 hours in 0.75^ silver nitrate; wash in distilled water 

 and harden in alcohol ; embed in celloidin. The cap- 

 illaries are brown-black on a pale yellow ground. 2. 

 For demonstration of lattice fibers : Harden for 2 days 

 in o. 5 r r chromic acid, treat for 3 days with 0.75$ sil- 

 ver nitrate, and further as for secretory capillaries. 

 The fibers are black. 3. For demonstration of cell 

 boundaries in the blastoderm of the bird. Fix for 2 

 or 3 hours in 3% nitric acid, to which \f solution of 

 silver nitrate has been added. Bolton's Method for 

 nerve tissue. Harden in 5 % formalin and mordant 

 with osmic acid, iron alum, or ammonium molybdate. 

 See further Pal's modification of Weigert's method, 

 Table of Stains (Illus. Diet.). Bordeaux R, a 

 general stain, acting on cytoplasm and chromatin. It 

 is used in \% solution. Born's Method for smooth 

 muscle: Isolate in potash lye, transfer to glycerol, and 

 add repeatedly 2 or 3 drops of glycerol acidified with 

 hydrochloric acid and of tincture of iodin, until the 

 brown color imparted by the latter reagent does not 

 fade. The iodin, which eventually fades, may be re- 

 placed by a carmin stain. Boston's Mixture for the 

 preservation of casts in urine : Liquid acidii arseniosi 

 (U.S. P.), 1 fluidounce; salicylic acid, % grain; 

 glycerin, 2 fluidrams. Dissolve by warming gently 

 and add "whole tears" of acacia to saturation. Let 

 the mixture settle, decant the supernatant liquid, and 

 add a drop of formalin. Place a drop of urine con- 

 taining casts on a slide, evaporate nearly to dryness, 

 add a drop of the preservative, mix the two with a deli- 



cate needle, apply a cover-glass, and when the mount 

 has hardened seal with cement. Bostroem's 

 Method for staining actinomyces in tissue sections : 

 Stain for from I to 3 hours in anilin gentian violet and 

 without washing transfer to Weigert's picrocarmin 

 (see staining Reagents) ; wash in water and extract in 

 alcohol until the sections are red-yellow. Bottcher's 

 Method for preparation of sperm crystals: Evaporate 

 a drop of spermatic fluid on a slide and stain with a 

 strong solution of iodin in solution of potassium iodid. 

 The crystals will be brown or violet. Charcot- Leyden 

 crystals stain yellow with iodin. Bouin's Liquid 

 for fixing tissues : I . Seventy-five volumes of a satu- 

 rated solution of picric acid, 25 volumes of formol, 5 

 volumes of glacial acetic acid. 2. Ten parts each of 

 formol and I % solution of platinum chlorid. 3. 

 Twenty parts each of I ft solution of platinum chlorid 

 and saturated solution of sublimate, 10 parts of formol, 

 and 3 parts of acetic or formic acid. Bowhill's 

 Method for the flagella of bacteria : Treat the prepa- 

 ration for 15 minutes with a mixture of 15 c.c. of a 

 saturated alcoholic solution of orcein, 10 c.c. of a 2of c 

 solution of tannin, and 30 c.c. of distilled water. 

 Wash and examine in water. Braddon's Method 

 for making blood-films : Accurately appose two cover- 

 glasses and seal three edges with vaselin or cement, 

 leaving open a very little of the edge opposite the un- 

 sealed one. Place the unsealed edge in contact with a 

 drop of blood, which will diffuse in. a thin even film 

 between the covers, and complete the sealing. Brass's 

 Formula, chromic acid and acetic acid each I part, 

 water 400 parts. Bremer's Method for diabetic 

 blood : Fix the films for 6 minutes in the oven at 135 

 C. Stain for 3 minutes with I <f ( solution of methyl- 

 blue, or with the Ehrlich-Biondi mixture. The yellow- 

 green reaction of the erythrocytes may also be obtained 

 by using eosin, congo red or biebrich scarlet in I </ c so- 

 lution. Bristol's Method for the regeneration of 

 reduced solutions of osmic acid (Os0 4 ): Contact with 

 organic matter reduces the tetroxid to the dioxid 

 (OsO,), which is regenerated by oxidizing with hydro- 

 gen dioxid. The reaction that takes place is expressed 

 in the following equation: OsO, -+- 2rl,0,= Os0 4 -4- 

 2H,0. Buchner's Method. I. for the cultivation 

 of anaerobic bacteria : Place the inoculated tubes, 

 with the cotton plug loosely inserted, in a vessel with 

 a capsule containing a mixture of pyrogallol and 

 liquor potassi, each I part, and water 10 parts; the 

 vessel should be closed with an air-tight cover. 2. 

 For staining spores : Treat the preparation for a half- 

 minute with concentrated sulfuric acid ; rinse in water 

 and stain in carbol-fuchsin. Bunge's Method for 

 the flagella of bacteria: Fix the film in the flame and 

 treat it with a mixture of tannin solution, 3 volumes, 

 and diluted liquor ferri sesquichlorate (1: 20), one 

 volume, containing carbol-fuchsin in the proportion of 



1 : 10. Bunge-Trantenroth Methcd for smegma 

 bacilli : Place the fresh cover-glass film for 3 hours in 

 absolute alcohol, then for 15 minutes in chromic acid; 

 carefully wash in water frequently changed. Stain for 



2 minutes in boiling carbol-fuchsin; decolor for 3 

 minutes in dilute sulfuric acid or for 2 minutes in pure 

 nitric acid. Counterstain in concentrated alcoholic so- 

 lution of methylene-blue, for about 5 minutes. The 

 smegma bacilli are blue, the tubercle bacilli red. 

 Busch's Method. I. For the myelin of nerves: 

 Place formalin material for 5 days in a solution of one 

 part osmic acid, 3 parts sodium iodate, 300 parts 

 water. 2. For tissues that have undergone fatty de- 

 generation : Fix for 2 days in 5^ formalin and then 

 in Flemming's liquid in the usual way. Treat the sec- 

 tions with 0.5^ chromic acid for 3 hours, then with 



