1()0 Connective Tissue (chap. 1!') 



smooth muscle — deep purplish red 

 epithelial cytoplasm — varies, red to green 

 collagen — clear green 



comments: 



This method gives excellent differentiation between smooth muscle, 

 collagen and fibrin. 



Koneff's Aniline Blue Stain (1936) 



fixation: any general fixative, preferably containing HgCU- 



solutions: 

 Iron alum: 



ferric ammonium sulfate 5.0 gm. 



distilled water 100.0 ml. 



Hematoxylins, see page 124. 

 Aniline blue solution: 



aniline blue WS, C.I. 42780 0.1 gm. 



oxalic acid 2.0 gm. 



phosphomolybdic acid 15.0 gm, 



distilled water 300.0 ml. 



procedure: 



1 . Deparaffinize and hydrate slides down to \vater, remox e HgCU. 



2. Mordant in iron alum: 10 minutes. 



3. Wash in distilled water: 10 ininutes, several changes. 



4. Stain in hematoxylin (Delafield's or the like): 5-15 minutes. This 

 may be progressive or regressive. 



5. Wash in running water: 5 minutes; rinse in distilled water. 



6. Stain in aniline blue stain: 10-15 minutes. Koneff stains over- 

 night. 



7. Rinse in distilled water; few dips. 



8. Dehydrate quickly through 50 and 70% alcohol. 



9. Dehydrate in 95% alcohol, 2 changes: 3 minutes each. 

 10. Finish dehydration in absolute alcohol; clear and mount. 



results: 



collagen, reticulum, mucus, hyaline cartilage — blue 

 nuclei, red corpuscles — violet or purple blue 

 cytoplasm, myelin sheaths, muscle fibers — brown 

 elastic fibers — reddish brown or red 



