DS 12.212-DS 12.221 DYE STAINS OF GENERAL APPLICATION 327 



12.212 Other Formulas 



12.212 Chatton 1920 eosin Y -light green 1915,59:21 



REAGENTS REQUIRED: A. 95% alc. 100, light green 1, eosin Y 2; B. 5% acetic acid in 



abs. alc. 

 PREPARATION OF a: Dissolve with occasional agitation over period of some days. Filter. 

 method: [sections with red nuclei] — » 95% alc. —* A,b mins. -^ B, till connective tissue 



clear green -^ balsam, via usual reagents 

 result: on arthropod material, for which the stain was designed, chitin is green on a 



red background. On vertebrate material the picture is similar to DS 12.32 Patay 1934. 



12.212 Kostowiecki 1932 orange G-anilin blue 23632, 49 :337 



formula: water 100, anilin blue 0.06, orange G 0.2, phosphomolybdic acid 1 

 preparation: Boil dyes with water 3 minutes. Add acid to hot sokition. Cool. Filter. 

 method: [sections with red nuclei] — > water -^ stain, till dark colored, l^ to 12 hrs. -^ 



water, rinse -^95% alc, 1 min. -^ balsam, via usual reagents 

 result: procartilage, light blue; cartilage, dark blue; muscle, orange; other connective 



tissues, blue green. 



12.212 Roux 1894 dahlia-methyl green 766,9:248 



formula: water 90, abs. alc. 20, dahlia violet 0.5, methyl green 0.5 

 preparation: Grind each dye separately in 10 abs. alc. Wash out each mortar with 

 50 water in small successive doses. Collect washings; leave 24 hours; filter. Mix 

 filtrates; leave 24 hours filter. 

 method: [red nuclei] -^ water — > stain, 5-15 mins. -^ blot — > abs. alc, till differentiated 



— * balsam, via usual reagents 

 note: The dahlia violet used by Roux may have been almost any mixture of pararo- 

 saniline derivatives. Cheap samples of gentian violet work admirably. The result is 

 an excellent polychrome counterstain wherever reproducibility of research results is 

 of less importance than classroom clarity of demonstration. The working solution 

 has been evaporated to dryness and sold as Roux's blue, (See DS 13.5 Bohm and 

 Oppel 1907.) 



12.212 Unna test. 1928 Hill anilin blue-orcein Gatenby and Cowdry 1928, 280 



reagents required: A. 0.1% acetic acid; B. water 50, abs. alc. 25, acetic acid 2.5, 



glycerol 10, orcein 0.5, anilin blue 0.5 

 preparation of B: Dissolve blue in water with gentle heat. Filter. Dissolve orcein in 



alc. and add to it the acid and glycerol. Add this mixture to the blue. 

 method: [red nuclei—* A, few minutes —> B, 1 to 10 hrs. -^ A, till differentiated —> 



balsam, via usual reagents 

 result: bone and elastic fibers red-brown against a blue background. 

 note: This reaction is more usually applied by the method of Pasini (1928) (DS 12.31). 



12.22 CONTRASTS FOR BLUE NUCLEI 



12.221 Formulas Containing Picric Acid 



12.221 Curtis 1905 picro-ponceau 1863, 17 :603 



formula: water 100, ponceau S 0.1, picric acid 1, acetic acid 0.04 



12.221 Fite 1939 picro-fuchsin 11284,25:743 



formula: water 100, picric acid 0.5, acid fuchsin 0.1 



12.221 van Gieson 1896 picro-fuchsin 23632, 13 :344 



formula: sat. sol. (circ. 1.2%) picric acid 100, acid fuchsin 0.05 

 method: [blue nuclei] —> water ^ stain, 2-10 mins. -^ water, quick rinse —> balsam, 



via usual reagents 

 note: a detailed description of the use of this stain is given under 12.20 above. 



12.221 Gnanamuthu 1931 picro-congo red 11360,51:401 



formula: sat. sol. {circ. 1.2%) picric acid 50, ammonia 50, Congo red 2 

 preparation: Add the ammonia to the picric solution. Dissolve the dye in mixture and 

 boil till no odor of ammonia is apparent. Cool. Add sufficient water to redissolve ppt. 

 formed on cooling. 



