Our Science - Tissue Array Research Program (TARP)
H&E Protocol For Array Slides On Tape Section Slides
The following suggested protocol was provided by
David E. Kleiner M.D., Ph.D
and Stephen M. Hewitt, M.D.,Ph.D.
- Staining the Instrumedic's acrylic coated slides used for tape sectioning.
- It may be used for general H&E's as well.
- This stain is programmed on our Leica Autostainer XL.
- Slides cut onto regular slides may be stained per routine.
| Step | Reagent | Time |
|---|---|---|
| 1 | Xylene | 2 minutes |
| 2 | Xylene | 2 minutes |
| 3 | Xylene | 2 minutes |
| 4 | 100% Ethanol | 1 minute |
| 5 | 100% Ethanol | 1 minute |
| 6 | 95% Ethanol | 1 minute |
| 7 | 95% Ethanol | 1 minute |
| 8 | Water | 2 minutes |
| 9 | Hematoxylin1 | 3 minutes |
| 10 | Water | 2 minutes |
| 11 | Bluing Agent2 | 30 seconds |
| 12 | Water | 30 seconds |
| 13 | Eosin Aqueous3 | 3 minutes |
| 14 | Water | 30 seconds |
| 15 | Oven 65° | 2 minutes |
| 16 | 100% Ethanol | 1 minute |
| 17 | 100% Ethanol | 1 minute |
| 18 | Xylene | 1 minute |
| 19 | Xylene | 1 minute |
| 20 | Xylene | 1 minute |
- We use Lerner-2 double strength hematoxylin, 0.36% hematoxylin.
- We use a LiCo3 0.044%, methanol 50%, bluing agent.
- It is essential aqueous eosin is used, or the acrylic adhesive on the slides will be stained.
We use Shandon Eosin Y Aqueous, 0.5% Eosin Y.
