310 Microorganisms (chap. 20) 



results: 



gTam-positive bacteria — deep violet 



oram-neoative bacteria — red 



erythrocytes — yellow 



nuclei — red 



cytoplasm — faint yellow 



comments: 



Glynn suggests treating sections with .OIN HCl before staining with 

 basic fiichsin. 



Lillie's Quick Method (1928) 



fixation: any good general fixative. 



solutions: 



Crystal violet solution: 



crystal violet, C.I. 42555 2.0 gm. 



957o ethyl alcohol 20.0 ml. 



ammonium oxalate, 1% (1 gm./lOO ml. water) 80.0 ml. 



Filter. Keeps well. 



Lugol's solution, see page 410. 



Safranin: 



safranin O, C.I. 50240 0.5 gm. 



distilled water 100.0 ml. 



procedure: 



1. Deparafhnize and hydrate slides to water. 



2. Stain in crystal violet solution: 30 seconds. 



3. Wash in running water: 3-5 minutes. 



4. Treat with Lugol's solution: 30 seconds. 



5. Wash in running water: 2-3 minutes. 



6. Flush with acetone from dropping bottle until no more color is 

 removed: 10-15 seconds. 



7. Wash in water. 



8. Cotuiterstain in safranin: 30 seconds. 



9. Rinse in distilled water. 



10. Dehydrate and differentiate in acetone. Some of red differentiates 

 out of cells leaving nuclei red and cytoplasm pink. 



11. Clear and mount. 



results: 



gram-positive bacteria — blue-black 

 gram-negative bacteria — red 



