SECTION TWO 



HAEMATOXYLIN (Heidenhain) - VAN GIESON STAIN 



A selective stain for collagen and connective tissue, 

 superior to Haematoxylin (Ehrlich) - Van Gieson 



Solutions required: 



A. Haematoxylin (Heidenhain) A. 



B. Haematoxylin (Heidenhain) B. 



C. Van Gieson stain (Picro fuchsin) 



Technique: 



1. Fix pieces of tissue in Bouin, Carnoy, Susa or io% formalin. 



2. Fix sections to sUdes ; dewax and take down to water in the 

 usual way, after removing mercurial precipitate if Susa has been 

 used as the fixative. 



3. Immerse in solution A for one half to one hour. 



Note: If a fixative other than Bouin, Carnoy, Susa or formalin 

 has been used it will be necessary to increase the time in solution 

 A and in solution B up to twelve hours or longer : the time varies 

 for different fixatives. 



4. Rinse in water. 



5. Stain in solution B for a time exactly equal to step 3. 



6. Rinse in water. 



7. Differentiate with solution A, controlling by examination 

 under the microscope, after the preparation has been rinsed briefly 

 in water. 



8. Wash gently in running water for about five minutes to 

 remove all traces of solution A (iron alum). 



9. Stain for three to five minutes in Van Gieson. 

 ID. Rinse for a few seconds in water. 



11. Examine, while still wet, under the microscope. 



12. Continue the staining with Van Gieson, or continue the 

 differentiation with water, whichever is necessary. 



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