STAINS 



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STAINS 



for IO seconds with 0.5^ solution of potassium per- 

 manganate, and wash in aicohol ; or, place the stained 

 sections in 1Z% solution of iron sulfate for 10 seconds, 

 or, in \of solution of iron chlorid. 7. For epithelia. 

 Stain sections for 10 minutes in neutral aqueous I °/ 

 solution of water blue ; wash, and stain for 10 min- 

 utes in 1 f solution of orcein. Or, overstain sections 

 of alcohol material in polychrome methylene-blue and 

 differentiate in Unna's glycerin-ether mixture (Griib- 

 ler) ; or, in a mixture of alcohol 10 parts, xylol 15 

 parts, anilin 25 parts, and transfer to xylol ; or, in a 

 mixture of xylol 30 parts, alcohol 20 parts, then trans- 

 fer to xylol and then to anilin containing alum to sat- 

 uration (agitated and filtered before using). 8. For 

 plasma-cells and mast-cells. Apply the methods for 

 epithelia. For the demonstration of bacteria in epi- 

 thelia see the methods for staining microorganisms. 

 9. For overcoming the decoloration of bacteria in the 

 process of dehydrating in alcohol. Transfer the sec- 

 tion from the decolorizing fluid to the slide, remove as 

 much as possible of the water by means of filter- 

 paper, and then heat the slide over flame until the 

 section is dry ; when cold mount in balsam. Unna- 

 Tanzer Stain. See Tamer's Orcein. Van Bene- 

 den-Neyt Method for the nuclear spindle and astro- 

 spheres of the ova of ascaris: Fix with acetic-acid 

 alcohol and stain with malachite green prepared by dis- 

 solving a little of the dye in glycerol diluted with 2 

 volumes of water. Van Ermenghem's Method 

 for the flagella of bacteria : Fix the film for a half- 

 hour at room-temperature or for 5 minutes at 6o° C, 

 in a mixture of one part of 2% osmic acid, 2 parts of 

 20% tannic acid, and 5 drops of glacial acetic acid. 

 Wash in water, then in alcohol, and treat with 0.5% 

 silver nitrate solution for a few seconds. Transfer 

 into a mixture of potassium acetate, 10 gm. , tannin, 3 

 gm., gallic acid, 5 gm., distilled water, 350 c.c, and 

 after a few seconds place again in the silver solution, 

 until this begins to blacken. Van Gieson's 

 Method for amyloid substance. See Table of Stains. 

 Result : amyloid, rose to brown-red. Vedeler's 

 Method for the "protozoon " of lipoma: Fix small 

 cubes of the tissue in a 5$ solution of mercuric 

 chlorid; extract the fat with ether (frequently renewed 

 for several weeks) ; harden in alcohol ; stain with 

 hematoxylin and eosin; embed in paraffin. Result: 

 lying in the empty fat-capsules are oval vacuolated 

 forms, resembling nuclei of endothelial cells undergo- 

 ing hyaline degeneration, and dark violet circular 

 bodies from 7 a to 16 fi in size, with a blue-black 

 limiting membrane and a nucleolated nucleus. Vial- 

 leton's Method for the blastoderm of the chick, be- 

 fore the appearance of the primitive streak : Open 

 the egg in salt solution, cut the blastoderm from the 

 yolk and put it on a slide; treat it with 1% silver 

 nitrate solution, wash, and put into 70% alcohol for 

 from 6 to 12 hours, in the dark. Stain with borax - 

 carmin and mount in balsam. Vincenzi's Method 

 for chromophilic granules: Tease fresh gray tissue of 

 the brain or spinal cord in normal salt solution, place 

 a drop of the emulsion on a slide, and stain with 

 methylene-blue. Cf. Turner's Method. Violet B, 

 a preparation of methyl-violet used in solution of I 

 gm. in 303 c.c. of 0.5% salt solution for staining 

 fresh tissues. It is a specific stain for the elements of 

 the vascular system. Potassium acetate may be used 

 as a mounting medium. Violet of Lauth. A name 

 for thionin. Vogel's Method for the study of the 

 origin and development of the connective tissue replac- 

 ing the fibrinous exudate after acute pneumonia : Stain 

 the sections for 24 hours in Tanzer's orcein, wash in 

 water and differentiate in acid alcohol ; wash, and stain 



for 15 minutes in Loffler's methylene-blue; bleach for 

 a few minutes in 70% alcohol. Von Rath's Mix- 

 ture for fixing tissues. I. Cold saturated solution of 

 picric acid 350 c.c, osmic acid 0.25 gm., and afte; 

 several hours add I c.c. of acetic acid. Fixing re- 

 quires from 15 minutes up to 48 hours, according to 

 the size of the object. Transfer from the fixing fluid 

 to 75% alcohol. 2. Mix 100 c.c. each of saturated 

 aqueous solutions of picric acid and mercuric chlorid ; 

 add 20 c.c. of 2% osmic acid. These mixtures are 

 recommended for fixing mitotic figures. 3. Cold satu- 

 rated solution of picric acid I part, hot saturated 

 solution of sublimate I part, and glacial acetic acid 

 \ c /o. Fix in this mixture for several hours and trans- 

 fer to alcohol. Wager's Method for staining the 

 yeast plant : Fix for 12 hours in sublimate or for 24 

 hours in 1:2: 300 iodin potassium iodid solution ; 

 wash in water, in 30^, 70%, and in methyl-alcohol. 

 Place a drop containing yeast cells on a slide, let the 

 alcohol evaporate and add a drop of water. When the 

 cells settle, drain and dry by evaporation. Add an- 

 other drop of water and stain with fuchsin and 

 methyl-green. Waldeyer's Method for the fixing 

 and decalcification of bone : Fix the fresh object in 

 chromic acid (i:6co); decalcify in a mixture of 

 chromic acid (I : 2Co) 100 c.c. and nitric acid 2 c.c. 

 Wash thoroughly and harden in alcohol. Wash- 

 burn's Medium for preserving fresh-water sponges 

 and other museum specimens : Mix 2 volumes of pure 

 glycerin and I volume of 3$ formalin. This mixture 

 is valuable because it does not extract color and per- 

 manently retains its transparency. Water Blue, an 

 acid dye resembling methyl blue and used in a concen- 

 trated aqueous solution, by Mann in conjunction with 

 eosin for staining ganglion-cells; by Mitrophanow asa 

 double stain with safranin. Stain chromosmium tis- 

 sue first in water-blue, for from-12 to 24 hours, then in 

 safranin for from 4 to 5 hours. Weigert's Method. 

 I. For neuroglia : Fix for 8 days in the following 

 mixture: dissolve 2.5 gm. chrome alum in 100 c.c. 

 water, by heat, and while hot add 5 parts each of 

 acetic acid and pulverized copper acetate, when cold 

 10 parts of formalin. Embed in celloidin. Treat the 

 sections for 10 minutes with 0.3% solution of potas- 

 sium permanganate, wash in water and reduce in the 

 following : 5 parts each of chromogen and formic acid 

 in 100 parts of water, to which after filtering add 10 

 parts of a \O c / solution of sodium sulfite. After 3 

 hours transfer to 5 % chromogen and after 24 hours 

 stain in the following: saturate hot 75^ alcohol with 

 methyl violot, decant when cold and to each ico c.c. 

 add 5 c - c - °f S7 P a q ueous solution of oxalic acid; 

 differentiate in a saturated solution of iodin in 5 J, so- 

 lution of potassium iodid; decolor in a mixture of 

 equal volumes of anilin and xylol, wash in xylol and 

 mount in balsam. 2. For elastin. see Weigert's Re- 

 sorcin-fttchsin. Weigert's Picrofuchsin. Warm 

 saturated picric acid solution, 150 c.c, saturated arid 

 fuchsin solution, 3 c.c. Weigert's Resorcin-fuch- 

 sin. Dissolve I gm. of basic fuchsin and 2 gm. of 

 resorcin in 200 c.c. water; heat to the boiling joint 

 and add 25 c.c. of liquor ferri sesquichlorati, P. G., 

 and boil for from 2 to 5 minutes, stining meanwhile. 

 Filter when cool, restore the precipitate on the filter to 

 the capsule, add 200 c.c. of 95', alcohol, ami boil. 

 When cold, filter, bring the filtrate up to 200 c.c. with 

 alcohol and add 4 c.c. of hydrochloric acid. Stain 

 sections for from 20 minutes to an hour and wash in 

 alcohol. (Avoid essential oils. ) This is a specific 

 stain for elastin. The elastic fibers are dark blue on a] 

 light ground. Nuclei may be afterstained in carmin. 

 Welcke's Method for flagella: Prepare a film from 



