CHAPTER XV. 173 



have not noted the proportions. SQUIRE (Methods and Formula, 

 p. 42) takes 1 grm. Saurefuchsin, 6 grins. Orange G in 60 c.c. of 

 alcohol and 240 c.c. of water. See also under " Connective tissues." 



295. EHRLICH-BIONDI Mixture (or EHRLICH-BIONDI-HEIDENHAIN 

 Mixture) (Pfluger's Arch,, xliii, 1888, p. 40). 



To 100 c.c. saturated aqueous solution of orange add with con- 

 tinual agitation 20 c.c. saturated aqueous solution of Saurefuchsin 

 (Acid Fuchsin) and 50 c.c. of a like solution of methyl green. 



(According to Krause (Arch. mik. Anat., xlii, 1893, p. 59), 100 parts of 

 water will dissolve about 20 of Saurefuchsin (Rubin S), 8 of orange G 

 and 8 of methyl green.) The solutions must be absolutely saturated, 

 which only happens after several days. 



Dilute the mixture with 60 to 100 volumes of water. The dilute 

 solution ought to redden if acetic acid be added to it ; and if a drop 

 be placed on blotting-paper it should form a spot bluish green in the 

 centre, orange at the periphery. If the orange zone is surrounded 

 by a broader red zone, the mixture contains too much fuchsin. 



According to M. HEIDENHAIN (" Ueber Kern u. Protoplasma," in 

 Festschr.f. Kolliker, 1892, p. 115) the orange to be used should be 

 ' Orange G," the Acid Fuchsin or Saurefuchsin should be '* Rubin 

 S ' (" Rubin ' is a synonym of Fuchsin) and the methyl green 

 should be " Methylgriin 00." And it is absolutely necessary that 

 these ingredients be those prepared under those names by the 

 Actienfabrikfiir Anilin-fabrikation in Berlin. They can be obtained 

 from Griibler & Hollborn, either separately, or as a mixture of the 

 three dyes in powder (which I do not recommend). 



The strong solutions directed to be taken readily precipitate on 

 being mixed. To avoid this it is recommended by SQUIRE (Methods 

 and Formulce, etc., p. 37) to dilute them before mixing. 



Other proportions for the mixture have been recommended by KRAUSE 

 (loc. cit. supra), viz. 4 c.c. of the Saurefuchsin solution, 7 of the orange G 

 and 8 of the methyl green ; the mixture to be diluted 50 to 100-fold with 

 water. THOME (Arch. mik. Anat., lii, 1898, p. 820) gives the proportions 

 2:5:8, and dilutes 100-fold. 



Stain sections (N.B. sections only) for six to twenty-four hours. 

 Dehydrate with alcohol, clear with xylol, and mount in xylol 

 balsam. 



In the intention of the observers who have elaborated this stain 

 it is a progressive stain, and not a regressive one. It does not require 

 any differentiation, and the sections should be got through the 

 alcohol into xylol as quickly as possible in order to avoid any extrac- 



