MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



Results: 



Lignified tissues are stained red, while unlignified tissues and 

 cytoplasm are blue-green. Nuclei, red. Chloroplasts, pink to red. 



SAFRANIN - ACID FUCHSIN 

 For spermatozoids, zoospores, motile gametes, etc. 



Solutions required: 



A. Osmic acid i% aqueous. 



B. Safranin i% aqueous. 



C. Acid Fuchsin i% aqueous. 



Technique: 



1 . Place a drop of an aqueous suspension of the organism on a 

 slide. 



2. Fix by holding the slide for about one minute over osmic 

 acid solution. 



3. Allow the preparation to dry. 



4. Stain for ten minutes to one hour in solution B. 



5. Wash in water. 



6. Remove excess water by draining and blotting the edges of 

 the slide with filter paper. 



7. Wash in 95% alcohol until only the nuclei retain the stain. 



8. Stain in the acid fuchsin solution for ten to twenty seconds. 



9. Wash rapidly in 70% followed by 90% alcohol. 



10. Wash rapidly with two changes of absolute alcohol. 



11. Clear in clove oil, followed by xylol. 



12. Mount in Canada balsam in xylol, Cristalite, Clearmount 

 or Emexel mountant. 



Results: 



Nuclei bright red. Cytoplasm bluish pink. 



Reference: (19 18), Steil. Bot. Gaz., LVX, 592. 



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