Phosphotungstic Acid-Hematoxylin (PTAH) Staining - Staining Protocol
Sample type required: Deparaffinized and rehydrated tissue section (3-5 microns) on positively (+) charged slides
Preferred fixative: Zenkers; however 10% neutral buffered formalin (NBF) may be used as well
Control: Skeletal or cardiac muscle
5 minutes (in heated solution)
Used as both a fixative and mordant.
Microwave solution for 45 seconds on high power before placing slides inside.
|Running water wash||5 minutes||Ensure that ALL traces of the reagent have been removed.|
|Lugol's iodine ||5 minutes|
|Running water wash||5 minutes||Ensure that ALL traces of the reagent have been removed. |
|5% hypo solution (sodium thiosulfate)||1 to 2 minutes|
|Running water wash ||10 minutes|
|0.25% potassium permanganate||5 minutes||Oxidizes|
|Running water wash ||5 minutes|
|5% oxalic acid ||Bleach until tissue is colorless|
|Running water wash||5 minutes|
|Distilled water ||3 changes||Discard solution after use.|
|PTAH staining solution||Overnight at room temperature|Post staining procedure:
Dehydrate quickly through two changes each of 95% and 100% alcohol, along with two changes of xylene, then cover-slip.
- Cross striations - Blue
- Nuclei - Blue
- Collagen - Red-brown
- Elastic fibers - Purplish