30 THE AMEFJCAN MONTHLY [February, 



Notice of New Methods— YII. 



By GEORGE C FREEBORN, M. D., 



INSTRUCTOR IN NORMAL HISTOLOGY, COLLBGE OF PHYSICIANS AND SURGBONS, NEW YORK. 



Microscopical Staining. Greisbach, H. — At the meeting of the 

 Anatomical Society lield at Wiirzburgh in May, i8SS. the author 

 demonstrated several methods of double, triple and quadruple staining. 

 The dyes used were in part anilines of the azo group, and part those 

 of other groups. They were all used in concentrated aqueous solu- 

 tions, either in combination or as single successive stains. The dyes 

 all bear careful washing in water and alcohol. The stained specimens 

 were cleared in anis oil and mounted in balsam. 



The combinations employed were as follows : 



DOUBLE STAINS. 



Metanil Yellow [Phenylamidobenzolmetasulphonate of soda] and 

 Azo Blue [Tetraazoditolylbetanaptholdisulphonate of soda] . — Prep- 

 aration : ala nasi of a child, alcohol hardening. 



The sections are stained in a mixture of equal parts of the two stain- 

 ing fluids for lo minutes, or for ten minutes in the yellow fluid and 

 then for 4 minutes in the blue. 



The epidermis, hair shaft, inner root-sheath, striated and smooth 

 muscle stain yellow ; the rete Malpighi, the outer root-sheaf, sebaceous 

 and sweat glands stain brownish-yellow ; connective tissue, elastic 

 fibres, and membrane of fat cells stain violet-blue ; hyaline cartilage 

 and nuclei do not stain. 



Metanil Yellow and Methyl Green. — Preparation : ala nasi of a 

 child, alcohol hardening. 



The sections are stained in a mixture of 5 c.c. of the yellow staining 

 fluid and 3 c.c. of the green. A crystaline precipitate forms which 

 does not interfere with the staining. The sections are allowed to re- 

 main in this fluid for 8 minutes or longer [^ of an hour], or they are 

 stained for 8 minutes in the yellow fluid and then for 1 minute in the 

 green . 



Epidermis, hair-shaft, inner root-sheath, striated and smooth muscle 

 stain yellow; the rete Malpighi, outer root-sheath hair-follicle, sweat 

 and sebaceous glands, and nuclei stain green ; hyaline cartilage and 

 cells stain green. 



Metanil Yellow and Crystal Violet [Hydrochloride of hexame- 

 thylpararoaniline]. — Preparation : ala nasi of child, alcohol hardening. 



Mix 7 c.c. of the yellow fluid with 3 c.c. of the violet. An amorphous 

 precipitate results which does not interfere with the staining. The 

 sections are stained in this mixture for 6 minutes, or they are stained for 

 10 minutes in the yellow fluid and then for 30 seconds in the violet. 



Epidermis, hair shaft, inner root-sheath, connective tissue, and elastic 

 fibres, the membrane of fat cells, and striated muscle stain yellow ; the 

 rete Malpiglii, the outer root-sheath, all glands, smooth muscle, and 

 cartilage with its cell nuclei stain violet. 



Metanil Yellow and Safranin. — Preparation : human lip, alcohol 

 hardening. 



Mix 6 c.c. of the yellow fluid with i c.c. ot satranin. An amorphous 

 precipitate forms. This mixture gives either with a long or short stain 

 more sharp pictures than the successive single staining. 



