STAINS 



500 



STAINS 



aqueous solution of ferric chloric! ; drain and dry and 

 stain for 3 minutes in a I J n aqueous solution of hema- 

 toxylin ; wash and differentiate in a 0.25^, solution of 

 ferric chlorid. Result : nuclei, dark blue ; fibrin, gray 

 to dark blue. In sublimate preparations the erythro- 

 cytes are greenish gray ; connective tissue, pale yellow. 

 5. For staining AmcebcE coli in tissues : Use alcohol 

 material and treat the sections for from 5 to 20 minutes 

 with saturated aqueous solution of thionin ; wash, and 

 differentiate for from 30 to 60 seconds in 2 Jo aque- 

 ous solution of oxalic acid ; wash, dehydrate, clear, 

 and mount in the usual way. Mallory- Wright 

 Method for staining tubercle bacilli : Stain lightly in 

 alum-hematoxylin, then for 2 or 3 minutes in steaming 

 hot carbol-fuchsin ; decolor for 30 seconds in acid alco- 

 hol. Manchot's Method for elastin : Stain sections 

 of alcohol or Miiller's fluid material for a half-minute 

 in saturated solution of fuchsin ; wash, and decolor in 

 acidulated syrup (10 c.c. of aqueous solution of sugar 

 of the consistence of glycerol plus 3 drops of sulfuric 

 acid). Mount in the nonacidulated syrup. Celloidin 

 sections must be fresh from the celloidin. Man- 

 naberg's Method for staining the malarial parasite : 

 Treat the air-dried film for 24 hours with a saturated 

 aqueous solution of picric acid diluted with an equal 

 volume of water and containing 3^ of glacial acetic 

 acid ; transfer to absolute alcohol ; stain with alum 

 hematoxylin; transfer to alcohol containing 25% of 

 hydrochloric acid and from this to alcohol containing 

 a little ammonia. Mann's Liquid. 1. For fixing 

 tissues : Ten parts each of saturated solutions of picric 

 acid and mercuric chlorid and 5 parts of formol. 2. 

 Equal parts of I Jo osmic acid and 5 J mercuric chlorid 

 in normal salt solution. Recommended for fixing the 

 tissue of the central nervous system. Mann's Method 

 for attaching sections to the slide : Shake I part of 

 white of egg with 10 parts of distilled water and filter 

 twice through the same paper. Apply to slides by 

 means of a glass rod ; let them drain and dry. Float 

 the sections on water warmed to 40 C. ; pass a slide 

 beneath them, arrange, lift them out, and subject 

 them for 5 minutes to 35 C. Manson's Method for 

 malarial blood : Take up a very small drop of blood 

 on a slip ( I y 2 X H m - ) °^ tnm d ssue -p a P er » about a 

 half inch from the end. When the blood has diffused 

 in a film, place the paper in contact with the slide or 

 cover-glass and draw it over the surface. For the 

 detection of crescents prepare a thick film ; dry ; fix 

 in absolute alcohol and dissolve out the hemoglobin 

 in very dilute acetic acid (2 or 3 drops in 30 c.c. of 

 water). Marchi's Method for the granules of tissue 

 cells undergoing fatty degeneration. See Staining of 

 Nerve Tissue, Table 0/ Stains (Ulus. Diet.). Mar- 

 choux's Stain for the parasite of malaria : Add 20 

 c.c. of a saturated solution of thionin in 50 J% alcohol 

 to 100 c.c. of 2 c / carbolic acid, and let the mixture 

 stand for a few days before using. Marina's Fluid : 

 Chromic acid 10 gm., formalin 5 c.c, g$fc alcohol 

 100 c.c. Stir until the acid is dissolved and let the 

 solution stand several hours before using. Marina's 

 Method. 1. For ganglion-cells : A modification of 

 Ileld's counterstain. Mix 3 c.c. of Nissl's methylene- 

 blue, 3 c.c. of 5% aqueous solution of acetone, and 30 

 drops of I Jo aqueous solution of erythrosin. Stain the 

 sections for 2 days and differentiate after Nissl. 2. 

 For ganglion-cells and medullated nerve-fibers : Fix 

 in Marina's fluid for 24 hours or longer, according to 

 the size of the object. Glue the tissue with syndeticon 

 on cork, treat for 2 hours with 95% alcohol, and cut 

 sections ; stain for 24 hours in Nissl's methylene-blue, 

 for 2 hours in erythrosin (see Ileld's Method), and 

 treat for from 12 to 24 hours with a mixture of equal 



parts of saturated solution of copper acetate and ot 1 J 

 lithium carbonate, plus enough ammonium hydroxid 

 to dissolve the precipitate; wash with distilled water 

 and stain for 24 hours in lithiated Weigert's hema- 

 toxylin at 35 C. ; differentiate after Weigert. Mar- 

 schalko's Method for plasma cells : Stain sections 

 of alcohol material in borax-methylene-blue or thionin 

 and differentiate in acidulated water or in "]oJ c alcohol 

 and dehydrate in absolute alcohol. Marzinowsky's 

 Method for the differential staining of human and 

 avian tubercle bacilli and lepra and smegma bacilli : 

 Stain films or sections from 3 to 8 minutes in carbol- 

 fuchsin diluted with 2 volumes of water ; wash, and 

 stain from 3 to 5 minutes in Loftier' s methylene-blue. 

 The avian tubercle bacillus stains red and is not de- 

 colorized by alcohol. The human tubercle bacillus 

 cannot be stained by this method. The lepra bacillus 

 stains red and is decolorized by alcohol. The smegma 

 bacillus stains red and with prolonged staining in 

 methylene-blue turns violet and finally blue. Maupas' 

 Method for the study of infusoria: Cultivate Para- 

 mecium caudatum or P. aurelia in a solution of boiled 

 flour frequently renewed. Place several individuals on 

 a slide in a moist chamber. Conjugation soon begins 

 and continues for about 12 hours. Fix in sublimate 

 solution and stain in methyl-green. Maurer's Stain 

 for blood: Solution A. — To al^ aqueous solution 

 of methylene-blue (med. puriss., Hochst, or Anilin- 

 blau, Merck) add 0.5% Na 2 CO s . Expose to the sun 

 for 2 or 3 days, or keep at room-temperature for 8 

 days. Add % J formalin to prevent formation of 

 mold. Solution B. — A o. I J aqueous solution of eosin 

 (Grubler, w. g. ). Maurer gives two methods of pro- 

 cedure : (1) With rapid; (2) with slow staining. 

 lie also indicates grades of intensity of staining. I. — 

 Rapid staining. The blood-film is placed at an angle 

 face down on a glass slide, one edge being elevated. 

 Solutions A and B are used in their full strength. I. 

 Grade of intensity of staining is obtained by mixing 2 

 parts of Solution A with 20-12 parts Solution B. 2. 

 Grade of intensity of staining is obtained by mixing 2 

 parts of solution A with 10-4 parts Solution B. 3. 

 Grade of intensity of staining is obtained by mixing 2 

 parts of Solution A with 3—2 parts Solution B. 4. 

 Grade of intensity of staining is obtained by mixing 2 

 parts of Solution A with 2-1 parts Solution B. As 

 the period of most intense staining lasts only a few 

 seconds after the mixture is made up, it is neces- 

 sary to stain immediately, and the staining need not be 

 carried on for more than a few minutes. II. — Slow 

 staining. I c.c. Solution A is mixed with 25 c.c. water. 

 1 c.c. Solution B is mixed with 25 c.c water. These 

 solutions are then mixed in a beaker. Films to be 

 stained are immersed in the stain immediately. I. 

 Grade of intensity of staining is obtained in 10 minutes. 

 2. Grade of intensity of staining is obtained in 20 

 minutes. 3 and 4. Grades of intensity of staining are 

 obtained in l / % hour to I hour at the longest. In 

 Grade I the nuclei of leukocytes stain blue or bluish 

 violet. Blood-platelets stain pale blue. Malarial para- 

 sites — cytoplasm stains pale blue; chromatin stains 

 ruby red. In Grade 2 nuclei of leukocytes stain violet 

 red. Chromatin of malarial parasite stains brilliant 

 red. In Grade 3, Grade 2, with Schiiffner's mottling 

 of infected red blood-corpuscles, in addition. In 

 Grade 4, Grade 3, and in addition in malarial parasites 

 the achromatic area about the chromatin stains faintly 

 red. Mayer's Carmalum and Indigo-carmin : 

 Disssolve o. 1 gm. of indigo-carmin in 50 c.c. of dis- 

 tilled water or of 5% alum solution ; add one volume 

 of indigo-carmin solution to 4 volumes of carmalum. 

 Mayer's Hemalum and Indigo-carmin : Add 



