STAINING, PRACTICAL AND THEORETICAL 



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. 



References : 



Foot, N. C. (1933)- 

 Masson, P. (1929). 



SAFRANIN - CRYSTAL VIOLET - FAST GREEN - 



ORANGE 2 



(Kalter's quadruple stain) 



Solutions 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. 



Technique: 



1. Any fixative may be employed, but if Bouin is chosen, remove 

 picric acid with a few drops of saturated lithium carbonate in the 

 70% alcohol of dehydration series. Should a fixative containing 

 mercuric chloride be chosen, then it will, of course, be necessary 

 to remove mercuric precipitate in the usual way. 



2. After fixation, wash, dehydrate, clear and embed in paraffin 

 wax. 



3. Cut sections no thicker than y-^fi and fix to slides, avoiding 

 the use of glycerine albumin, which will cloud the stain. 



4. De-wax with xylol ; then pass through absolute, 90% and 

 70% alcohol to water. 



5. Stain in the safranin solution for twenty-four hours. 



6. Rinse with water. 



7. Stain with the crystal violet for one to two minutes. 



396 



