Sample type required: Deparaffinized and rehydrated tissue section (3-5 microns) on positively (+) charged slides
Preferred fixative: 10% neutral buffered formalin (NBF)
Control: Aorta, artery or skin
Verhoeff's elastic staining solution
| 1 hour | Make fresh; save solution until staining is complete. Ferric chloride is required as a mordant in this (hematoxylin-based) solution. |
| Running water wash | Until solution drains clear |
|
2% ferric chloride solution | Differentiate until black fibers are crisp and well defined and background is gray
| Review slides microscopically to ensure differentiation is complete. Due to varying elastic fiber content, the control slide and patient slide should be checked individually and microscopically
to determine accurate differentiation. If differentiation is not complete, return to solution for 30 seconds and repeat. If over-differentiation occurs, return slide(s) to Verhoff solution.
|
| Running water wash | Until solution drains clear |
|
5% sodium thiosulfate | 1 minute | Removes iodine |
| Running water wash | 5 minutes |
|
Van Gieson's solution
| 5 minutes | Counterstain |
Post staining procedure: Tissue section should be rinsed well in distilled water, dehydrated through graded alcohols(beginning with 95%). Follow with two changes of xylene and then coverslip.
Expected results:
- Elastic fibers and nuclei - Black
- Collagen - Red
- Other tissue elements - Yellow
