SECTION TWO 



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



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

 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. 



10. 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. 



13. Drain and draw off excess water by means of a filter paper 

 applied carefully to the edges of the section, but do not allow the 

 preparation to dry completely. 



14. Dehydrate with absolute alcohol only. 



15. Clear in xylol. 



16. Mount in D.P.X., Cristalite or Clearmount. 



Results: 



Nuclei of cells: dark brown to black. Collagen fibres: bright 

 red. Erythrocytes, muscle, epithelia and other tissues: yellow. 



Note: Van Gieson stain fades if mounted in Canada balsam, but 

 fading can be avoided by the use of D.P.X., or Cristalite or Clear- 

 mount. 



129 



