MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



2. Pieces, which must not be thicker than 0-5 cm., are immersed 

 in Solution A for twenty-four hoiirs. 



3. Transfer to Solution B for twenty-four hours. 



4. Transfer to Solution C for forty-eight hours ; then wash in 

 water for twenty-four hours. 



5. Dehydrate in the usual manner. 



6. Clear in beechwood creosote for twenty-four hours ; then in 

 benzol for twenty-four hours. 



7. Embed in a saturated solution of paraffin wax in benzol at 

 room temperature; then successively in saturated solutions of 

 paraffin wax in benzol at 38° C, 42° C. and 45° C. so that pure 

 paraffin wax only is used for the final embedding. 



8. Mount sections on slides: bring down to distilled water, 

 mordant sections on slides with Solution D for twenty-four hours, 

 then wash thoroughly in water. 



9. Stain for two hours with Solution E ; then rinse in tap water. 



10. Flood slides with Solution F and warm gently until vapour 

 is given off; or stain at room temperature for 24 hours. 



11. Rinse in 1% acetic acid; then dry by blotting carefully. 



12. Pass through absolute alcohol; then differentiate for about 

 ten minutes in beechwood creosote; dry by blotting carefully, 

 wash with xylol, and mount. 



Vital staining of nervous tissue in small vertebrates 



Solution required: 

 AHzarin Red, S, 2% aqueous 



Technique: 



1. Paraffin sections are brought down to distilled water by the 

 usual method. 



2. Stain twenty-four hours in 2% aqueous Alizarin Red, S. 



3. Differentiate thirty to sixty seconds in distilled water to 

 which has been added three drops 1% calcium acetate per 10 ml. 



4. Dehydrate : clear and mount. 



Note: This is a general stain which also demonstrates Nissl 

 bodies as well as other details. 



S8 



