MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



C. Gram's iodine solution. 



D. Carmalum. 



Technique: 



1. Thin pieces of fresh tissue (not more than 3 mm. thick) are 

 fixed for three to five hours in formalin-saUne solution. 



2. Make frozen sections and collect them in distilled water. 



3. Immerse in a mixture consisting of equal volumes of Solu- 

 tions A and B (Note: This mixture must be made and filtered 

 immediately before use) for about five minutes until the sections 

 turn blue. 



4. Rinse sections rapidly in distilled water. 



5. Immerse in Gram's iodine solution until the sections turn 

 brown. 



6. Transfer to distilled water to which two drops of lithium car- 

 bonate 0*5% aqueous have been added for each 100 ml. of distilled 

 water, for a quarter to twenty-four hours until the sections have 

 regained their blue colour. 



7. Wash in distilled water then counterstain in carmalum for 

 two to five minutes. 



8. Mount in glycerine jelly or Apathy's medium or in Aqua- 

 mount. 



Results: 



Oxidase granules are stained blue while nuclei are pink. Some- 

 times fat is stained also. 



NAPHTHOL BLUE BLACK - HAEMATOXYLIN - BRILLIANT 



PURPURIN AZOFUCHSIN 



For collagen, reticulum, smooth muscle, etc. 



Solutions required: 



A. Weigert's Haematoxylin A. 



B. Weigert's Haematoxylin B. 



C. Brilliant purpurin R. (C.I. No. 454) 

 in 1% acetic acid . . . . . . 30 ml. 



Azofuchsin 1% in 1% acetic acid. . 20 ml. 



168 



