0-5 gm. 

 100 ml. 



STAINING, PRACTICAL AND THEORETICAL 



CELESTIN BLUE - ORCEIN - LIGHT GREEN 

 For the study of skin lesions, etc. 



Solutions required: 



A. Rubens - Duval Orcein: 



Orcein o-i gm. 



Alcohol 70% 100 ml. 



Nitric acid, cone 2 ml. 



B. Celestin Blue 

 Iron alum 5% 



Shake the dye with the iron alum solution in a flask; 

 then boil for three minutes. 



Allow to cool ; filter; then add: 



Sulphuric acid, concentrated . . 2 ml. 



Glycerine . . . . • • • • ^4 ^^ 



C. Eosin yellowish aqueous 1% . . 2 volumes 

 Gallic acid 0-5% aqueous . . i volume 



D. Phosphomolybdic acid 1% aqueous. 



E. Masson's Light Green: 



Light green 2% aqueous . . . . 100 ml. 

 Glacial acetic acid 2 ml. 



Technique: 



1. Fix tissues in Zenker or Bouin; if the latter is used then 

 picric acid must be removed by washing de-waxed and dehydrated 

 sections on slides with saturated Uthium carbonate solution. If 

 Zenker is employed ; then mercuric precipitate must be removed 

 after fixation. 



2. Wash; dehydrate; clear; embed in paraffin wax as usual. 



3. Fix sections to slides ; remove paraffin wax with xylol. 



4. Pass through descending grades of alcohol down to distilled 

 water in the usual manner. 



5. Stain for one half to two hours in Solution A, in a stoppered 

 grooved staining jar in the incubator or for twenty-four hours at 

 room temperature. 



6. Rinse well with distilled water. 



166 



