SECTION TWO 



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 (Na2HP04). 



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. 



Results: 



Nuclei are stained blue. Cytoplasm, varying shades of red. 

 Muscle, pink. Elastic fibres, pink. Collagen, yellow. 



Improved differentiation of most cells and tissues is obtained by 

 employing this method in place of haematoxylin and eosin. 



SAFRANIN - CRYSTAL VIOLET - FAST GREEN - 



ORANGE 2 



(S. S. Kalter's " Quadruple Stain for Animal Tissues ".) Abstract from jf. 

 Lab. & Clin. Med., 28, 995-7, 1943. 



This technique, which is a development of Flemming's 

 triple stain, is particularly useful for histology students 



Solution required: 



A. Safranin, O . . . . . . 0-2 gm. 



Formalin (40% formaldehyde) . . 4 ml. 



Sodium acetate . . . . . . 0-5 gm. 



Alcohol 50% . . . . . . 100 ml. 



B. Crystal violet 0-5% aqueous. 



C. Fast green F.C.F. - Orange G 



saturated in clove oil 



D. Orange 2 saturated in clove oil. 



195 



