STAINING, PRACTICAL AND THEORETICAL 



8. Wash and blue in tap water. 



9. Remove excess water by draining and blotting very carefully. 



10. Stain with solution B for ten to fifteen minutes, or longer 

 (up to twenty-four hours). 



11. Rinse briefly in distilled water, then drain and carefully 

 remove excess water by blotting. 



12. Differentiate in solution D (clove-toluol), controlling under 

 the microscope, for about ten to fifteen minutes. 



13. Rinse with two changes of toluol. 



14. Mount in permount-toluene or in Clearmount or Cristalite. 



Results: 



Zymogen granules: dark violet. Parietal cells, scarlet, and 

 nuclei, blue. The parietal cells are distinctly contrasted from the 

 pepsinogen cells. 



Reference: Cambel, P. & Sgouris, J. (195 1). 



BISMARCK BROWN - METHYL GREEN 



For mucin, cartilage, and goblet cells in embryonic tissue, 



trachea and intestine 



Solutions required: 



A. Bismarck brown 1% aqueous. 



B. Methyl green 0-5% aqueous. 



Technique: 



Tissues are fixed in Bouin or Zenker and embedded in paraffin 

 wax. 



1. Sections are brought down to distilled water; then stained 

 five to ten minutes in Solution A. 



2. Wash with 95% alcohol. 



3. Stain with Solution B until the preparation appears dark 

 green to the naked eye. 



148 



