I'l.ASiMA STAINS WITH OOAL-TAIi DYKS. 189 



of the acidified stain added, dehydrates, clears with xylol, 

 and mounts in xylol-balsani. Connective-tissue red, elastin 

 and all other elements yellow. 



WEIGERT (Zeit. iciss. Mik., 1904, p. 3) adds 10 parts of 1 

 per cent. Saurefuchsin to 100 of saturated picric acid. 



See also MOLLEE, op. cit., xv, 1898, p. 172. 



This stain is generally used as a contrast stain to follow 

 lisematoxylin. APATHY (JBehrenis' Tabdlen, 3rd ed., p. 129) 

 takes for this purpose 1 grin, of Saurefuchsin in 500 c.c. of 

 saturated solution of pier ate of ammonia. 



WILHELMI (Fauna Flora Golf. Neapel, xxii, 1909, p. 18) 

 takes 0'2 grm. Saurefuchsin, 0'8 grm. picrate of ammonia, 

 10 grm. absolute alcohol, and 89 grin, water. 



E. and T. SAVINI (Zed. wiss. Nik., xxvi, 1909, p. 31) use 

 a formula due to BENDA. Ninety-five volumes of saturated 

 solution of picrate of ammonia are mixed with 5 volumes of 

 1 per cent, solution of Saurefuchsin. For use, two to four 

 drops of saturated solution of picric acid are added to 10 c.c. 

 of the mixture. This neither overstains nor attacks the 

 primary stain. 



300. FLEMMING'S Orange Method (Arch. mik. Anat., xxxvn, 1891, 

 pp. 249 and 685). Stain sections of Flemning or Hermann material in 

 strong alcoholic safranin solution diluted with anilin water ( 286) ; 

 differentiate in absolute alcohol, containing at most OT per cent, of 

 hydrochloric acid, until hardly any more colour comes away ; stain for 

 one to three hours in gentian violet ( 287) ; wash for a short time in 

 distilled water ; treat with concentrated, or at least fairly strong, 

 aqueous solution of orange Gr. After at most a few minutes, whilst 

 pale violet clouds are still being given off from the sections on 

 agitation, bring them into absolute alcohol until hardly any more colour 

 comes away, clear in clove or bergamot oil, and mount in damar or 

 balsam before the last pale clouds of colour have ceased to come away. 

 The orange must be orange G. 



WINIWARTER and SAINMONT (Zeit. wiss. Mile., xxv, 1908, p. 157, 

 and Arch. BioL, xxiv, 1909, p. 15) stain for 24 hours in the gentian, 

 wash out after the orange for 2 to 3 hours in 100 c.c. absolute alcohol 

 with three to four drops of HC1, and differentiate finally with oil of 

 cloves. 



This is not a triple stain in the sense of giving three different 

 colours in the result ; it is a nuclear and plasmatic stain in mixed tones; 

 the orange, apparently, combines with the gentian to form a " neutral " 

 dye, soluble in excess of the orange ( 203) which thus differentiates the 

 stain. 



