STAINS 



501 



STAINS 



one volume of a 0.05 % aqueous solution of mdigo-car 

 min to 4 volumes of hemalum. Meek's Method 

 for elastin : Stain sections of alcohol material in a 

 solution of orcein, 3 gm., in loo c.c. of absolute alcohol 

 and 40 c.c. of hydrogen dioxid ; after 3 or 4 minutes 

 differentiate in absolute alcohol and hydrogen dioxid 

 100:40. Melnikow-Raswedenkow's Fluid. 1. 

 For the preservation of macroscopic objects : Fix the 

 object for from I to 4 days in the following mixture : 

 sodium acetate 3 parts, potassium chlqrid 0.5 parts, 

 formalin 10 parts, water 100 parts ; transfer it into 

 strong alcohol and from this into a solution of potas- 

 sium acetate 30 parts, glycerol 60 parts, water 100 

 parts. 2. For the preservation of bile pigment : 10 V 

 solution of formalin containing I c /o of hydrochinone. 

 Melnikow-Raswedenkow's Method for the study 

 of the '• echinococcus " of tyrolean jaundice: Fix 

 the organ for 24 hours in 4$ formaldehyd, harden in 

 alcohol, and embed in celloidin. Stain for 30 minutes 

 in Weigert's resorcin fuchsin ; wash ; differentiate in 

 90 r ' c alcohol for 2 minutes ; dip into weak solution 

 of lithium carbonate; rinse and stain in alum hema- 

 toxylin and van Gieson's picrofuchsin. Mercier's 

 Method for blood in tissue sections : Fix in Zenker's 

 fluid ; after 24 hours harden in alcohol and subse- 

 quently treat with iodin alcohol. Metachromatic 

 Stain, one which imparts different colors to different 

 tissues. Methyl Blue, Methyl Water Blue. See 

 Water Blue. Meyer's Method for staining nerve- 

 tissue with methylene-blue : At intervals of 15 minutes 

 inject subcutaneously 2 c.c. of a saturated aqueous 

 solution of methylene-blue BX at body-temperature. 

 Fix the tissue for 24 hours in Betlie's fluid. Michaelis' 

 Method. I. For staining fat : Treat frozen sections, 

 hardened in formalin, with a saturated solution of 

 scharlach R in 70% alcohol, for 15 or 30 minutes, and 

 mount in glycerol or levulose. 2. For the nuclei of 

 leukocytes : Prepare I <& solution of pure methylene- 

 blue and eosin in fresh, nonalkaline, distilled water ; 

 (a) mix 20 c.c. of the methylene-blue solution with 

 20 c.c. of alcohol and (b) 12 c.c. of the eosin solution 

 with 28 c.c. of acetone. At the time of using mix I 

 c.c. each of a and b and keep the mixture covered. 

 Fix the film of blood for 24 hours in absolute alcohol 

 and submerge it, film side down, in the stain. The 

 time for staining is from one half to 10 minutes and 

 must be tested for each preparation. The action of 

 the dye should be stopped when the film turns from 

 blue to red. Michaelis' Stain for blood : Solution 

 A. — The polychrome methylene-blue solution. To 

 200 c.c. of a I c 'c aqueous solution of methylene-blue, 



add IO c.c. NaOH solution. Boil the mixture for 



10 



15 minutes. After cooling neutralize with 10 c.c. 



N 

 H 2 S0 4 solution. Solution B. — A 0. I ^ aqueous 



solution of eosin. To 2 c.c. of Solution A, add 10 c.c. 

 of Solution B. Stain blood-films in this mixture tor 

 15 minutes. Wash rapidly in running water. Blood- 

 films must be thin ; those in which the cells remain 

 spherical do not take the violet chromatin stain. Fix 

 blood-films for I hour in absolute alcohol. Minot's 

 Method for embryonic epidermis and developing 

 hairs: Macerate the embryo for several days in o. 6'V 

 salt solution containing o. 1 ^ of thymol. Miquel's 

 Medium for the cultivation of bacteria : Prepare a 

 solution of 10 parts of sodium chlorid and I part of 

 potassium carbonate in 1000 c.c. of water and add 4 

 parts of gelatin. Mitrophanow's Method. A modifi- 

 cation of Weigert's for medullated fibers : I. Mordant 

 photoxylin sections for 24 hours at 40 C. in a 

 mixture of equal parts of saturated aqueous solution 



of copper acetate and 90 f r alcohol, stain for IO 

 minutes in Kultschitzky's hematoxylin and differen- 

 tiate with Weigert's ferricyanid. 2. Or, after the 

 copper bath stain for 10 minutes in acid hema- 

 toxylin (1 gm. in 400 c.c. of absolute alcohol plus 

 4 c.c. of acetic acid), differentiate in 0.25^ potassium 

 cyanid in 45% alcohol, and when the photoxylin is 

 decolored put into the same with the addition of I J t 

 solution of red prussiate of potash. Moll's Method 

 for the study of embryonic cartilage : Fix in alcohol and 

 stain thin celloidin sections for from 6 to 24 hours in 

 Tanzer's orcein, wash in 90^ alcohol until the celloidin 

 is nearly bleached, dehydrate in absolute alcohol, and 

 clear in oil of origanum. Result : preformed hyaline 

 cartilage blue- violet, all else brownish-red. Moller's 

 Liquid for fixing vegetal organisms : A saturated 

 solution of iodin in 1^ solution of potassium iodid. 

 Money's Method for bacieria in tissues : Stain the 

 sections in picrocarmin ; then in gentian-violet or 

 methylene-blue, adding a few drops of formalin and 

 heating until it steams. Wash in water and decolor- 

 ize in 90% alcohol. Morgan's Method for produc- 

 ing abnormal cleavage in the eggs of sea-urchins : 

 Place them in sea-water to which 2j£ or less of 

 sodium or magnesium chlorid has been added and 

 after a short time restore them to normal sea-water. 

 Cleavage occurs in unfertilized eggs and spindles, cen- 

 trosomes, and chromosomes appear in abnormal posi- 

 tions. Morse's Method for pathologic tissues: Fix 

 in any medium, preferably in formalin or sublimate. 

 Dissolve I gm. of kresylviolet (Leonhard) in a mix- 

 ture of 80 c.c. of 5jt aqueous solution of phenol and 

 20 c.c. of 95 5£ ethyl alcohol, and filter. Stain the 

 sections for from I to 5 minutes, wash in distilled 

 water, dehydrate in anilin-xylol (2: 1), clear in xylol 

 and mount in balsam. Result : nuclei, blood-plate- 

 lets, and basophil granules violet or rose-pink ; cyto- 

 plasm pale blue or pale green ; intercellular substance 

 of connective tissue dull rose pink ; cartilage reddish 

 violet ; elastic fibers sky blue ; axis-cylinders and cell- 

 body of nerve-cells purple ; plasmodium malaria; dull 

 pink ; colloid substance deep indigo blue ; amyloid 

 substance ruby red ; mucin bright rose pink ; the 

 so-called cancer parasite rose pink ; the granules of 

 mast-cells (staining 10 seconds and differentiating in 

 alcohol) fuchsin red. Muchematein, a specific stain 

 for mucin. I. Pulverize 0.2 gm. of hematin with a 

 few drops of glycerol and then add o. I gm. of alumi- 

 num chlorid, 40 c.c. of glycerol, 60 c.c. of water. 2. 

 Dissolve 0.2 gm. of hematin and o. I gm. of aluminum 

 chlorid in 100 c.c. of 7°/£ alcohol. Two drops of 

 nitric acid may be added. Mucicarmin, a specific 

 stain for mucin. Rub I gm. of carmin in a mortar 

 with 0.5 gm. of aluminum chlorid and 2 c.c. of dis- 

 tilled water; heat for 2 minutes, until the light red 

 color has become dark ; stir and add a little 5°% a l c °- 

 hol; when dissolved make up to 100 c.c. with 50^ 

 alcohol and after 24 hours filter. For use dilute ten- 

 fold with water or with 50% alcohol. Miiller's 

 Method. I. For blood in "sections" : Float the 

 dry cover-glass preparation for one minute on a very 

 thin solution of celloidin ; drain ; when dry strip off 

 the "section" of blood in celloidin and stain. 2. For 

 staining tubercle bacilli : Stain with carbol-fuchsin in 

 the usual way and decolor with potassium perman- 

 ganate and oxalic acid after Pal's modification of 

 Weigert's method for medullated nerves. See Table 

 of Stains (Illus. Diet.). 3. For the study of secre- 

 tory capillaries in the gastric glands: Treat the tissue 

 for 24 hours in a mixture of 3.5% potassium bichro- 

 mate, 40 parts, and formalin. 10 parts ; then for I or 2 

 davs with the bichromate alone. Wash in water and 



