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STAINS 



alcohol g parts, and clear in cajeput oil. 2. Stain 

 overnight in concentrated solution of toluidin blue, 

 wash in water, differentiate in alcohol. Erythrosin 

 may be used after the thionin and the toluidin blue. 

 Lepkowsky's Method for the study of the blood- 

 supply of the teeth : Inject with Berlin blue, harden 

 in 5j' f formalin, decalcify in io^i nitric acid, fre- 

 quently renewed, and make celloidin sections. Lewin- 

 son's Method for staining adipose tissue: Fix in 

 Muller's fluid and embed in celloidin. Stain the sec- 

 tions for 12 hours in hematoxylin (I gm., in enough 

 absolute alcohol to dissolve it, and 50 c.c. of 2^ acetic 

 acid); wash in water and treat for 15 minutes with 1 v 

 solution of potassium permanganate ; wash, and treat 

 for 5 minutes with 2' c oxalic acid; wash, and counter- 

 stain for 24 hours in an ammonia solution of carmin; 

 differentiate for 2 minutes in acid alcohol and stain for 

 1 minute in a saturated alcoholic solution of picric acid. 

 Lewis's Method for tracing medullated nerves : Place 

 a small piece of brain tissue in 10 times its volume of 

 I fo osmic acid. Renew the solution after 2 days and 

 4 days. In 5 or 10 days wash in water and harden in 

 alcohol. Treat the sections with a drop of ammonia. 

 Mount in soluble glass. Lightfoot's Anilin Black, 

 a preparation similar to anilin blue-black. Cf. Xoir 

 colin. Loeb's Method for producing artificial par- 

 thenogenesis: Place the unfertilized eggs of sea-urchins 

 in sea-water containing magnesium chlorid in the pro- 

 portion of 5000 {\°n MgCl) to 5000 c.c. of water. 

 After 2 hours restore them to normal sea-water. The 

 eggs form normal gastruke and plutei. Loffler's 

 Stain for flagella: Mix 10 c.c. of 20% solution of 

 tannin, 5 c.c. of saturated solution of ferrous sulfate, 

 and I c.c. of aqueous or alcoholic solution of fuchsin, 

 methyl-violet, or " Wollschwarz. " For typhoid 

 bacilli add 1 c.c. of 1 r c soludon of soda; for Bacillus 

 subtilis add 30 drops; for bacilli of malignant edema 

 36 drops. For cholera bacilli add one drop of sulfuric 

 acid to the soda solution; for Spirillum rubrum 9 drops. 

 Lowit's Method for fibrin: In a fresh cover-glass 

 film let the blood coagulate, then wash off the erythro- 

 cytes with 0.6% salt solution, and apply Weigert's 

 method (q. v.). See Examination of the Blood, 

 Table of Stains (Illus. Diet). Lubarsch's Method. 

 I. For glycogen in tissue cells : Apply Weigert's 

 method for fibrin, allowing the iodin solution to act but 

 for a very short time. See Examination of the Blood, 

 Table of Stains (Illus. Diet. ). 2. For tumors : Harden 

 very small cubes of the tissue for from a half to three- 

 fourths of an hour in alcohol several times renewed. 

 Place in anilin at 50 C. for a half to one hour and for 

 the same time in xylol, renewed until it does not 

 become yellow. Infiltrate with paraffin once renewed 

 for from I to 2 hours ; then embed. By this method 

 stained sections can be made in a few hours. Luith- 

 len and Sorgo's Method for ganglion-cells: Stain 

 celloidin sections of material hardened in alcohol or in 

 Orth's or Muller's fluid for 24 hours in polychrome 

 methylene-blue heated until it steams; wash for 24 

 hours in distilled water several times renewed, differ- 

 entiate in Unna's glycerin-ether mixture (Griibler); 

 absolute alcohol, origanum oil, balsam. Granules and 

 nucleoli of ganglion-cells and nuclei of glia-cells 

 violet ; connective tissue and axis-cylinders blue to 

 colorless ; medullary sheaths sometimes red-violet. 

 Lutschke's Stain : Ten c.c. of a 20 c / c solution of 

 tannin, 5 c.c. of a cold saturated solution of ferrous 

 acetate, 1 c.c. of a saturated alcoholic solution of 

 fuchsin. Lysol for the examination of fresh tissues : 

 It may be used in 10^ solution or in the following 

 mixtures : (a) lysol 10 parts, alcohol 30 parts, water 

 60 parts ; (b) lysol 10 parts, water 50 parts ; glycerol 



10 parts, alcohol 30 parts. McCrorie's Method for 

 flagella : Stain the cover-glass preparation in warmed 

 mixture of equal parts of a saturated solution of night 

 blue, a IO% solution of tannin, and a lo' /C solution 

 of alum. Magdala Red, a chromatin stain. See 

 Xaphlhalene Red under Pigments (Illus. Diet.). 

 Malachowski's Stain for blood : Stain in an aqueous 

 solution of eosin {percentage not stated, time not 

 stated). Then stain in "a very dilute aqueous solution 

 of borax-methylene-blue " (percentage not stated, 

 no time stated). Staining is performed rapidly by ap- 

 plying heat ; this, however, gives very uneven results. 

 \\ hen carried on in the cold, the stain is uniform, but 

 may require 24 hours. Blood-films are fixed in abso- 

 lute alcohol (time not stated). Malassez's Method 

 for staining bacteria in the blood : Prepare films on 

 cover glasses and dry them without heat ; wash in 

 distilled water or Ranviers alcohol and fix in chromic 

 or in osmic acid ; wash and stain. Mall's Method. 

 I. For the demonstration of noncollagenous reticu- 

 lated tissue, in the spleen, lymph-glands, mucous 

 membranes, liver, kidneys, and lungs : Digest sec- 

 tions with pancreatin, shake in a test-tube with water, 

 spread on a slide and dry by evaporation ; then treat 

 with a drop of picric acid (10 gm., dissolved in alco- 

 hol, 150 c.c, and water, 3C0 c.c.) and again dry by 

 evaporation. Stain for a half-hour with acid fuch- 

 sin (10 gm. , dissolve in absolute alcohol, 33 c.c. 

 and water, 66 c.c), treat for a few seconds with the 

 picric acid solution ; dehydrate in alcohol. 2. For 

 the demonstration of the "membranes" of elastic 

 fibers: Heat to boiling in strong hydrochloric acid 

 and pour acid and fibers into cold water. The 

 "membranes" may be isolated by boiling in 5^ 

 or \o c /o potash lye, also by treatment with pepsin, 

 which destroys everything but the sheath. Mallory's 

 Hematoxylin : Dissolve o.l gm. hematoxylin in a 

 little hot water and when cool add to IOO c.c. of lf c 

 phosphotungstic acid. Mallory's Method. 1. For 

 neuroglia: Fix for 4 days in 10^ formalin, then for 

 4 days in a saturated solution of picric acid ; after this 

 mordant for 4 days in 5 </ ( soludon of ammonium bi- 

 chromate at 37 C. Stain the sections for 2 minutes 

 in I f c aqueous solution of acid fuchsin, rinse, and 

 treat for 2 minutes with 1 <^ f aqueous solution of phos- 

 phomolybdic acid ; wash in 2 changes of water and 

 stain for 2 minutes in a mixture of water-soluble anilin 

 blue 0.5 gm., orange G 2 gm., oxalic acid 2 gm., and 

 water 100 c.c ; wash in water and dehydrate in alco- 

 hol. Result: connective tissue blue; neuroglia deep 

 red ; ganglion-cells and axis-cylinders light red. 2. 

 For neuroglia : Fix the tissues after the method given 

 in No. I, and treat the sections for 15 minutes with a 

 0.5 aqueous solution of potassium permanganate and 

 after washing for the same time with 1 r f solution of 

 oxalic acid ; wash, and stain in hematoxylin prepared 

 by dissolving o. I gm. of the dye in a little hot water 

 and when cool adding water up to 80 c.c, 20 c.c. of 

 io'r aqueous solution of phosphotungstic acid, and 

 last 0.2^ of hydrogen dioxid. Wash in water, de- 

 hydrate in alcohol, clear in oil of origanum, and mount 

 in balsam. Nuclei, neuroglia, and fibrin blue ; axis- 

 cylinders and ganglia-cells pale-pink; connective tissue 

 deep-pink. 3. For connective tissue : Fix in Zenker's 

 fluid or sublimate and stain the sections for 2 minutes 

 in O.I r f aqueous solution of acid fuchsin. For further 

 treatment see No. I. Result: fibrous tissue, mucus, 

 amyloid and hyaline substances, blue ; nuclei, cyto- 

 plasm, elastin, fibrin, neuroglia, and axis-cylinders, 

 red ; erythrocytes and myelin sheaths, yellow. 4. 

 For nuclei and fibrin : Stain sections of tissue fixed in 

 any medium except formaldehyd for 3 minutes in XO'fc 



