164 Connective Tissue (chap. 13) 



6. Wash in running water: 10 minutes. 



7. Rinse in 70% alcohol. 



8. Stain in resorcin solution: 16 hours. 



9. Wash in running water: 10 minutes; rinse in distilled water. 



10. Stain in Weigert's hematoxylin: 15 miniues. 



11. Wash in running water: 10 minutes; rinse in distilled water. 



12. Stain in woodstain-acid fuchsin: 5 minutes. (During its staining 

 action this sokition differentiates the hematoxylin.) 



13. Rinse in 0.5% aqueous acetic acid (0.5 ml./99.5 ml. water). 



14. Differentiate in phosphotungstic acid: 10 to 20 minutes, or until 

 collagen is pale pink and groiuid substance bluish. 



15. Rinse in 0.5% aqueous acetic acid. 



16. Dehydrate thorotighly in 3 changes of absolute alcohol. Use fresh 

 and good-grade alcohol or the tissue will not take on the collagen 

 stain which follows. 



17. Stain in saffron: 5 to 15 minutes. Use an airtight staining jar, 

 preferably screw-cap type of coplin jar, to prevent hydration of 

 saffron solution. 



18. Transfer throtigh 3 changes of fresh, good-grade absolute alcohol. 



19. Clear and moiuit. 



results: 



nuclei^ — blue to black 



cytoplasm — red 



elastic fibers — dark purple to black 



collagen and some reticular fibers — yellow to greenish yello^v 



ground substance (acid mucopolysaccharides) and some reticular 



fibers— blue to bluish oreen 

 fibrinoid — intense red 



comments: 



1. Movat says: "It is important for these staining methods to prepare 

 solutions with HCl of specific gravity 1:124 (the offizielle Salzaure) and 

 not concentrated HCl as stated in some textbooks. It is prepared by 

 adding 50 cc. of distilled water to 100 cc. of concentrated HCl (specific 

 gravity 1.19)." 



2. Movat includes other modifications of his method using orange G 

 and diphenyl fast red, but seems to favor the above procedtire. He calls 

 his method a demonstration of all connective tissue elements in a single 

 section. 



