STAINING, PRACTICAL AND THEORETICAL 



Carmine . . . , . . , . i gm. 



Ammonium chloride . . . . 2 gm. 



Boil for a few minutes ; allow to cool ; then make up 

 to the original volume and add : 



Strong ammonia solution . . 6 ml. 



Filter before use. 



B. Solution A . . . . • • 15 mi- 

 Pure methyl alcohol . . . . 12-5 ml. 

 Strong ammonia solution (sp. gr. 



o-88o) . . . . . . . . 2 ml. 



Distilled water .. .. .. 5-5 ml. 



C. Methylene Blue 1% in absolute 



alcohol. 



D. Picric acid, saturated aqueous . . 27 ml. 

 Sodium sulphate saturated 



aqueous . . . . • • 3 rnl* 



Technique: 



1. Fix thin slices of the material in absolute alcohol. 



2. Immerse for one-and-a-half to two hours in each of three 

 changes of acetone. 



3. Transfer to a mixture of equal volumes of acetone and benzol 

 for half an hour. 



4. Immerse in pure benzol for one half to one hour ; then embed 

 in paraffin wax. 



5. Fix sections to slides and de-wax with xylol. 



6. Pass through absolute, followed by 90% alcohol. 



7. Immerse in Solution B in a grooved staining jar for five 

 minutes, rocking gently, but continuously, during the period of 

 staining. 



8. Rinse thoroughly with absolute alcohol. 



9. Stain for half a minute in the methylene blue solution. 



10. Rinse with absolute alcohol. 



11. Stain for fifteen to thirty seconds in Solution D keeping 

 the slides in motion by rocking. 



12. Dehydrate thoroughly with absolute alcohol ; clear in xylol 

 and mount in balsam or cristalite. 



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