STAINING, PRACTICAL AND THEORETICAL 



Technique: 



1. Tissues are fixed in Bouin, Regaud, Zenker or formalin, and 

 embedded in paraffin wax. 



2. Mordant sections on slides with 5% iron alum previously 

 heated to 45° C. 



3. Wash with tap water; stain for five minutes in Regaud's 

 haematoxylin ; then rinse with 95% alcohol. 



4. Differentiate with picric alcohol; then wash with running 

 tap water. 



5. Stain for five minutes in violamine R-acid fuchsin; then 

 wash with distilled water. 



6. Differentiate for five minutes in 1% phosphomolybdic acid; 

 then without rinsing: 



7. Flood the slide with solution D and leave for five to ten 

 minutes. 



8. Rinse with distilled water; then return to 1% phospho- 

 molybdic acid for five minutes. 



9. Leave in 1% acetic acid for five minutes. 



10. Dehydrate in 95% alcohol, followed by absolute alcohol; 

 clear in xylol and mount. 



Results: 



Nuclei: black. Argentaffin granules: black or red. Cytoplasm: 

 vermilion. Collagen : green. Neuroglia fibrils : vermilion. Mucus : 

 green. Keratin: vermilion. Intercellular fibrils: vermilion. Golgi 

 apparatus : clear. 



Reference: Lillie, R. D. (1945). 



TRICHROME STAIN-PVA FIXATIVE 



For confirmation of Amoebiasis 



Solutions required: 



A. PVA-Fixative : 



Glacial acetic acid . . . . . . 5 ml. 



Glycerin . . . . . . . . 1*5 ml. 



Schaudinn's fixative .. .. 93-5 ml. 



Polyvinyl alcohol (PVA) . . . . 5 gm. 



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