Verhoeff-Van Gieson (VVG) Staining - Staining Protocol
Sample type required: Deparaffinized and rehydrated tissue section (3-5 microns) on positively (+) charged slides
Preferred fixative: 10% neutral buffered formalin (NBF)
Control: Artery or skin
Verhoeff (iron) hematoxylin
| 30 minutes | Make fresh; save solution until staining is complete. The ferric chloride in this particular hematoxylin is required as a mordant. |
| Running water wash | Until solution drains clear |
|
2% ferric chloride solution | Differentiate until black fibers are well defined and background is gray
| Review slides microscopically to ensure differentiation is complete. If differentiation is not complete, return to solution for 30 seconds and repeat.
|
| Running water wash | Until solution drains clear |
|
5% hypo (sodium thiosulfate) | 1 minute | Removes iodine |
| Running water wash | 1 minute |
|
Van Gieson's solution
| 5 minutes | Counterstain |
Post staining procedure: Tissue section should be dehydrated with 95% and absolute alcohols followed by two changes of xylene and then coverslip.
Expected results:
- Elastic fibers and nuclei - Black
- Collagen - Red
- Other tissue elements - Yellow
