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STAINS 



stration of iron in hematopoietic tissues: Fix the bone- 

 marrow of iron-fed rabbits for 24 hours in 70^ alcohol 

 containing 5 % of a solution of ammonium sulfate ; 

 transfer to absolute alcohol containing a few drops of 

 ammonium sulfate. Stain the paraffin sections after 

 Stieda (see Stieda's Method). 2. Fix in absolute 

 alcohol, treat the sections for an hour with solution of 

 ammonium sulfate, wash in distilled water, and mount 

 in glycerol. Cf. the methods of Hall and Zalewski. 

 Hoffmann's Method. 1. For the blastoderm of the 

 chick: Fix in lC/ nitric acid for 10 minutes and wash 

 in a 2% solution of alum. Fixation in nitric acid facili- 

 tates the isolation of the blastoderm ; in order to pre- 

 vent the curling of the edge during hardening it is 

 advised to spread the object on the convex surface of a 

 watch-glass. 2. For the orientation of small, opaque 

 objects in celloidin : After embedding, and hardening 

 in So% alcohol, treat for a time with 90% alcohol. 

 The consistency of the mass will then be such that the 

 object can be placed in the desired position. Treat 

 with xylol until hard and clear. Homberger's 

 Method for staining gonococci : Stain in the hanging 

 drop with a very dilute (1 : 10,000) aqueous solution 

 of kresyl violet. The gonococci take a reddish-violet 

 hue, while other microorganisms are faint blue or un- 

 stained. Honsell's Method for smegma bacilli : 

 Stain films for 2 minutes in boiling carbol-fuchsin ; 

 wash, dry, and treat for IO minutes with a mixture of 

 hydrochloric acid and absolute alcohol in the propor- 

 tion of 3 : 100. Wash, and stain in concentrated alco- 

 holic solution of methylene-blue diluted with an equal 

 volume of water. Huber's Method for the nerves 

 of the intracranial bloodvessels : Anesthetize the animal 

 and inject through the carotid, cerebral ward, enough 

 ifc methylene-blue in normal salt solution to tinge the 

 eye and ear of the same side. After a half-hour remove 

 the brain and cervical cord and expose to the air until 

 stained. Cut out bits of the cortex with curved scissors 

 and crush under a cover-glass until the gray substance 

 is pressed away from the pia. Ikeda's Method. See 

 Japanese Method. Intravitam Stain, one that will 

 act upon living material. Inversion, of Rawitz, a 

 process in which, under the influence of a mordant, 

 a basic anilin dye behaves as a plasma or acid 

 dye. Iodin-alcohol : Alcohol 90%; to which 

 enough tincture of iodin is added to impart the 

 color of port-wine. Cf. Zenker'' 5 Fluid. Iodin 

 Reaction. See the method of Ehrlich and of 

 Hofbauer. Iron Carmin Method: Stain in sections 

 in carmin for several hours, wash in dilute acetic acid, 

 and treat with I % ammoniated iron citrate until the 

 tissue becomes black ; wash for several hours in dis- 

 tilled water. Cf. von Wellheini s Stain. Israel's 

 Method for coloring actinomyces in sections: Stain 

 for several hours in a saturated solution of orcein in 

 water acidified with acetic acid. Jacottet-Sadowsky 

 Method for ganglion-cells : Harden pieces of the spinal 

 cord for from 2 to 4 days in 10% formalin ; transfer to 

 g$% alcohol, and after 48 hours to absolute alcohol. 

 Cut without embedding and stain I or 2 minutes in 

 carbol-fuchsin ; treat with acetic acid and then with 

 absolute alcohol. Sadowsky used a 5% solution of 

 methylene-blue instead of fuchsin. Jander's Method 

 for removing pigment from tissues. Fix in any suitable 

 medium, wash in water (if the object has been in alco- 

 hol), and treat for from 12 to 48 hours in a mixture,of 

 70 parts of 1 f solution of chromic acid, 3 parts of 

 potassium nitrate and 200 parts of water. The tissue 

 may be treated in bulk or in sections. Jaos' Medium 

 for the culture of diphtheria bacilli: Mix 50 c.c. of 

 normal sodium hydroxid solution, 150 c.c. of distilled 

 water, and 300 c.c. of blood-serum and heat over a 



water-bath for 2 or 3 hours at from 6o° to 70 C. and 

 then sterilize in steam for 45 minutes. Add 500 c.c. 

 of peptonized bouillon and 20 gm. of agar ; filter while 

 hot and sterilize for 15 minutes at ioo° or no° C. 

 and pour into petri dishes. The bacilli develop in 

 from 5 to 12 hours and, the medium being transparent, 

 the culture can be examined under the microscope 

 with a magnification of 60 to 70 diameters. Japanese 

 Method for mounting serial sections: Spread on a 

 slide as thin a film as possible of Mayer's albumen and 

 over this a little water ; arrange the sections on the 

 slide and cautiously warm over a spirit-lamp. When 

 the sections have spread out mop up the water and dry 

 at 35 C. Cf. Hentitguy? $ Method (2). Jelinek's 

 Method for washing objects fixed in picric acid : Use 

 alcohol to which a few drops of a saturated aqueous 

 solution of lithium carbonate have been added. This 

 makes the alcohol turbid, which becomes clear and 

 yellow in proportion to the extraction of the picric 

 acid. Add the carbonate from time to time, until the 

 object is entirely decolored. Jenner's Stain for blood. 

 Preparation of the neutral stain : In an open beaker 

 mix equal parts of 1.2 or 1.25^ aqueous solution of 

 eosin (Griibler), 1 % aqueous solution of methylene-blue 

 med. (Griibler). Let stand for 24 hours. Filter. Dry 

 the precipitate obtained. Wash the precipitate with 

 distilled water and dry again. The staining solution: 

 For use dissolve 0.5 gm. of the precipitate in loo c.c. 

 pure methyl alcohol (Merck "for analysis"). Jenner 

 gives no particular method of fixation. Staining : 

 Stain in the solution for 1 to 3 minutes, covering with 

 a watch-crystal. Pour off stain quickly and rinse in 

 water till film is pink (5 to 10 seconds). Staining re- 

 action: Leukocytes — nuclei stain blue; granules, neu- 

 trophil stain red ; granules, basophil stain dark 

 violet; granules, eosinophil brilliant crimson. Ma- 

 larial parasites, bacteria, and filaria, blue. Jensen's 

 Medium for the study of living infusoria: Dissolve 3 

 gm. of gelatin in IOO c.c. of water, by heat. Mix a 

 drop of this with a drop of the water containing the 

 organisms. The addition of a drop of very dilute solu- 

 tion of hematoxylin or of methyl green or other anilin 

 dyes will stain intra vitam. Joannovics' Method 

 for the study of plasma cells in pathologic processes. 

 Harden the tissue in formalin or in graded alcohols. 

 Stain the sections for 20 minutes in polychrome 

 methylene-blue and wash in water for 24 hours. Treat 

 with glycerin-ether (Griibler) until a cloud of color 

 appears. Dehydrate in 95% and absolute alcohol and 

 clear in origanum oil and then in xylol. Johne's 

 Method for staining the capsules of bacteria: Stain 

 the cover-glass preparations in a warmed 2 C / V solution 

 of gentian- violet, rinse, and differentiate for from 10 to 

 20 seconds in 2f c acetic acid ; wash and mount in 

 water. Jolly's Method of staining bone-marrow : 

 Place a slide gently on the fresh marrow ; fix the film 

 so obtained in the vapor of osmic acid or by treating 

 with strong liquid of Flemming for 15 minutes; wash 

 in water for 15 minutes and bleach for a second in 

 iodin and alcohol (l:lco); wash in alcohol, then in 

 water and overs tain in a solution of eosin I part, alco 

 hoi 20 parts, glycerol and water each 50 parts; decolor 

 in alcohol and stain in hematein I part, alcohol 25 

 parte, 5% solution of ammonia alum 2CO parts; water, 

 alcohol, carbol-xylol, balsam. Julinsburger's Method 

 for ganglion-cells : Slain sections of formalin material 

 for from a half to three-fourths of a minute in warm 

 1 solution of neutral red. Kaiser's Bismarck 

 Brown for staining kinetic nuclei: Prepare a saturated 

 solution of bismarck brown in bo', boiling alcohol 

 and in this stain lor 48 hours at 6o° C. Extract in 

 ih> , alcohol containing 2</ c of hydrochloric acid or 



