SECTION TWO 



SAFFRON - ERYTHROSIN 



For connective tissue 



Note: Saffron is the dried stigmata of crocus sativus, and should 

 not be confused with safranin, which is an anihne dye. 



Solutions required: 



A. Saffron . . . . . . . . 2 gm. 



Distilled water . . . . . . lOO ml. 



Boil gently for an hour; allow to cool; then filter; 

 add I ml. of 40% formaldehyde and i ml. of 5% 

 tannic acid to the filtrate. 



Note: Saffron solution deteriorates after a few 

 weeks, and it is best to prepare the solution in small 

 quantities, as required. 



B. Delafield or Ehrlich haematoxylin, 



C. Erythrosin, 1% aqueous. 



Technique: 



1. Fix small pieces of tissue in Bouin, Zenker-formaldehyde or 

 in mercuric-formaldehyde. 



2. Wash; dehydrate; embed. 



3. Sections are stained for five to ten minutes with Delafield or 

 Ehrlich haematoxylin; rinse in water. 



4. Blue in tap water in the usual manner or in 1% sodium phos- 

 phate (NagHPO^). 



5. Stain for two to five minutes in 1% aqueous erythrosin. 



6. Rinse quickly with water. 



7. Differentiate with 70% alcohol for a few seconds, controlling 

 under the microscope, until the collagen fibres are nearly colour- 

 less. 



8. Rinse in water; stain for five minutes in saffron solution 

 prepared as above; rinse with water. 



9. Wash rapidly first with 70% alcohol then with absolute alcohol ; 

 clear in xylol and mount. 



395 



