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hello i bought ab150683(PTAH staining kit) few weeks ago
this protocol is needed microwave for paraffin embedding tissue slide
can i following this protocol using frozen section? A frozen tissue are to weak when exposed heat or
how can i procedure?
Asked on May 20 2014
We can recommend to use charged slides for frozen section as thse should prevent the section from coming off the slide.
It is also possible to adapt the protocol to the use of a waterbatch instead of a microwave.
Heated Water Bath
1. Deparaffinize sections if necessary and hydrate to distilled water.
2. Pour Zinc Chloride Solution (10%) into plastic staining jar and set in 60° C. water bath for 10 minutes to equilibrate temperature.
3. Place slide in warmed Zinc Chloride Solution (10%) and incubate for 20 minutes at 60°C.
4. During step 3, pour Ferric Ammonium Sulfate Aqueous Solution into a second plastic staining jar and set in 60° C. water bath for 10 minutes to equilibrate temperature.
5. Rinse slide in running tap water for 1 minute.
6. Rinse in distilled water for 1 minute.
7. Place slide in warmed Ferric Ammonium Sulfate Aqueous Solution and incubate for 5 minutes at 60°C.
8. During step 7, pour Phosphotungstic Acid Hematoxylin Solution into a third plastic staining jar and set in 60°C. water bath for 10 minutes to equilibrate temperature.
9. Rinse slide in running tap water for 2 minutes.
10. Rinse in distilled water for 1 minute.
11. Place slide in warmed Phosphotungstic Acid Hematoxylin Solution and incubate for 60 minutes at 60°C.
12. Differentiate section in 95% Reagent Alcohol. Check section using microscope for proper differentiation.
Note: Graded alcohols will remove some stain.
13. Dehydrate in 3 changes of Absolute Alcohol.
14. Clear in 3 changes of fresh Xylene or Xylene Substitute, and mount in synthetic resin.
Anja HoffmannAbcam Scientific Support
Answered on May 20 2014