MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



GBEMSA - WRIGHT STAIN 



A permanent stain for differentiating the structures, 

 particularly Nissl bodies and cytons, of the spinal cord 



Solution required: 



Wright's stain . . . . • • 5 volumes 



Giemsa stain . . . . . . i volume 



Technique: 



1. Material should be fixed in neutral formalin io%. 



2. Wash, dehydrate, clear, and embed in paraffin wax in the 

 usual manner. 



3. Fix sections to slides; de-wax; pass through the usual 

 descending grades of alcohol, down to distilled water. 



4. Flood the sections with a measured volume of the above 

 staining solution and allow it to act for two minutes. 



5. Add an equal volume of distilled water and mix with stain 

 by rocking the slides gently. Allow this diluted stain to act for 

 two minutes. 



6. Pour off excess stain and immerse the slides in fresh dis- 

 tilled water for one minute. 



7. Transfer immediately into 80% alcohol and leave therein for 

 fifteen seconds. 



8. Dehydrate rapidly in 95% and absolute alcohol. 



9. Clear in xylol and mount. 



Results: 



Cytons and Nissl granules are stained deep blue. Nuclei of 

 blood-vessel structures and neuroglia are light blue. Elastic fibres 

 of blood vessels, deep blue. Erythrocytes, pink. Neuroglia fibres, 

 light red. 



Note: The proportion of the Giemsa stain regulates the inten- 

 sity of the cyton stain. 



Reference: Hanburg, L. (1935), Science, 81, 364-5. I 



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