MEDICAL AND BIOLOGICAL STAINING TECHNIQUES 



WATER BLUE - ORCEIN - SAFRANIN 

 For demonstrating epithelial fibres 



Solutions required: 



A. Water blue . . . . . . i gm. 



Orcein . . . . . . . . 0-75 gm. 



Glycerine . . . . . . . . 20 ml. 



Absolute alcohol . . . . 50 ml. 



Acetic acid 5% . . . . . . 100 ml. 



B. Eosin 1-25% alcoholic. 



C. Hydroquinone 1% aqueous. 



D. Safranin O, aqueous 1%. 



E. Potassium dichromate 0-5%. 



Technique: 



1. Specimens of skin are fixed in 10% formalin and embedded 

 either in paraffin wax or in Celloidin. 



2. Bring sections down to distilled water; then stain for ten 

 minutes in a mixture consisting of: 



Solution A . . . . . . 10 ml. 



Solution B . . . . • • 3 ^^ 



Solution C . . . . • • 3 rnl. 



3. Wash well in distilled water. 



4. Stain for ten minutes in the safranin solution. 



5. Wash thoroughly in distilled water. | 



6. Immerse in 0-5% potassium dichromate solution from ten 

 to thirty minutes. 



7. Wash in distilled water; dehydrate in absolute alcohol ; then 

 clear in oil of bergamot. 



8. Examine under the microscope; then if necessary differ- 

 entiate alternatively with absolute alcohol and oil of bergamot 

 until the depth of the safranin stain has been reduced. 



9. Mount in balsam or in Cristalite. 



Results: 



Epithelial fibres are stained red, while the nuclei are pale violet ; 



220 



