SECTION THREE 



5. Clear in xylol. 



6. Embed in paraffin wax. 



7. Cut sections and mount on slides. 



8. De-wax with xylol. 



9. Mount in Canada balsam in xylol. 



Note: For whole mounts, after clearing in xylol (Stage 5), 

 mount in balsam. 



Results: 



Nuclei are stained deep red, while cytoplasm is pink. 



CHLORAZOL AZURINE 



A simple double stain, non-fading, and particularly suitable 



for elementary class work 



Solutions required: 



A. Formaldehyde 40%. . .. .. 5 ml. 



Acetic acid 50% . . . . . . 14 ml. 



Absolute alcohol . . . . • . 63 ml. 



Distilled water . . . . . . 20 ml. 



B. Magnesium sulphate, crystals, 6% aqueous. 



C. Chlorazol azurine, saturated aqueous. 



D. Equal volumes of Solutions B and C. 



Heat the mixture to 80° C. and stir well for five or ten minutes, 

 taking care that this temperature is not exceeded by more than a 

 few degrees. 



Technique: 



1 . Sections of the material, fresh from the field, are transferred 

 to solution A. Alternatively material may be stored in this fixative 

 for several days before sectioning. 



2. Take sections down to water. 



3. Immerse sections in solution D (the solution should be 

 shaken well immediately before use) in a jar or tube overnight, or 

 for at least eight hours. 



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